Hepatitis B DNA test. Hepatitis B virus DNA, qualitative determination. What do the results mean?

Timely diagnosis of hepatitis can improve the prognosis of the disease and reduce the risk of such severe complications as cancer and cirrhosis of the liver. Early diagnosis is extremely important, if only because it allows timely determination of indications for antiviral therapy and carrying out immunoprophylaxis.

At the present stage There are 2 types of blood tests that are used to diagnose hepatitis:

quantitative and qualitative tests for DNA hepatitis virus.

In this article we will talk in detail about both research methods, find out how and when to take tests for hepatitis, what is a test for DNA, antibodies and a mysterious marker hbsag, and what the results of these tests may be.

Immunological diagnostic methods

There are many immunological diagnostic methods, but they do not have a clear classification, which can cause confusion. One way or another, each of them is based on the enzyme immunoassay method, or briefly - ELISA.

A little theory about the ELISA method

Structure of hepatitis C

Antibodies bind to antigens and enter into a complex immunochemical reaction with them, which can be detected in various ways. That's the point ELISA. For example, if a reagent with antibodies to hbsag, then when adding blood hbsag In a positive patient, antigen-antibody complexes are formed, by which the disease is judged.

What marker should you take a blood test for if you suspect hepatitis B?

Thus, laboratory diagnosis of viral hepatitis should be carried out only as prescribed by a doctor, who selects the scope of the study based on the results of examination and questioning of the patient. Attempts to diagnose hepatitis using laboratory tests on your own have no practical meaning and can lead to erroneous conclusions about your health.

Laboratory diagnosis of hepatitis B should be carried out only as prescribed by a doctor.

Australian antigen (HBV surface antigen) test

Virological diagnosis of hepatitis B

Virological diagnosis involves the isolation and identification of the virus. The method is used to detect the genetic material of the hepatitis virus in the blood. PCR –RT- polymerase chain reaction in real time. Using sophisticated equipment PCR allows you to detect and recognize HBV DNA(hepatitis virus) even if it is contained in small quantities. In this case, both qualitative and quantitative assessment of the result is possible.

Qualitative determination of hepatitis B virus (HBV) DNA

confirmation of a positive test by method ELISA ;
suspicion of infection with questionable results ELISA ;
mixed infection;
chronic liver diseases of unknown cause;
control of antiviral therapy.

For analysis, blood is taken from a vein. No special preparation is required for the procedure, although it is recommended to refrain from smoking 30 minutes before the test.

Quantitative determination of hepatitis B virus DNA

Viral load is an indicator that determines the number of copies of the hepatitis virus per milliliter of blood, and LDK- this is the range of values at which this indicator can be accurately determined. LDK depends on the capabilities of diagnostic equipment, and approximately ranges from 750 copies per ml to 100 million. Below and above these values, accurate viral load assessment is not possible.

Quantitative research on DNA solves a number of important problems:

monitoring the effectiveness of treatment;
detection of mutant strains of the virus;
determination of indications HTP ;
monitoring infection activity;
determining the prognosis of the disease.

Primary diagnosis of hepatitis B: answers to questions

In what cases is it worth getting tested “just like that”, without waiting for the manifestations of the disease?

After unprotected sexual intercourse, it is necessary to undergo comprehensive tests for HIV and hepatitis

In fact, such situations practically never occur. In theory test must be taken after each case where infection may have occurred:

after unprotected sexual intercourse;

after any contact with someone else's blood.

In practice, in both cases it is necessary to undergo complex tests for HIV and hepatitis, and in the first - another row STD why you need to visit a doctor. All people who, by their profession, are associated with a risk of infection undergo annual screening diagnostics and receive vaccination.

However, anyone can get tested for hepatitis at any time if they think they might have become infected. There is no such thing as too much vigilance, although this will not have any effect on the course of acute hepatitis, and for emergency preventive measures, if they are carried out, it is of greater importance HBV- source status and the presence of post-vaccination immunity in the victim.

Is it possible to do a rapid analysis at home?

Today, relatively inexpensive test systems for quick diagnosis are available on pharmacy shelves. HBV. The attitude of specialists towards these tests is ambiguous. On the one hand, it is a simple, fairly reliable and accessible screening tool that can be used periodically, for example, by members of the patient’s family. On the other hand, people who are at risk in any case undergo regular examinations and are vaccinated. One way or another, given that many people are not very willing to go to clinics and laboratories, home rapid tests can help in the early diagnosis of hepatitis.

According to manufacturers, the sensitivity of rapid rapid tests is 97-99%, and their cost in a pharmacy is around 170-250 rubles.

The most well-known tests in Russia include:

ImmunoChrom-HBsAg-Express (Med-express-diagnostics LLC, Russia);

iSCREEN-Hep (InTec PRODUCTS Inc., China);

New Vision Diagnostics (InTec PRODUCTS Inc., China).

All of the tests listed work the same. To carry out the analysis, you need to prick your finger with a scarifier, take a few drops of blood with a pipette and transfer an immunochromatographic strip, in the test zone of which there are antibodies to HbsAg. If the blood contains the Australian antigen, the latter reacts with the antibodies, causing a colored line to appear in the diagnostic window. In addition, a second strip always appears in the window, which serves as an indicator that the test is working. Thus, if a person is a carrier HbsAg, then the test will show two stripes.

Could there be an error in testing for hepatitis B using ELISA and PCR methods?

No one is immune from errors, but the vast majority of false-positive test results for hepatitis are caused by the human factor - violation of procedure technology, saving of reagents, defective or insufficiently high-quality consumables, intra-device contamination (contamination), material contamination, etc. Today we can reliably say that neither pregnancy, nor previous vaccination, nor any concomitant diseases affect the results of the analysis for the Australian antigen and PCR test for HBsAg

HBV is a complex DNA virus and belongs to the Hepadnaviridae family, genus Orthohepadnavirus.

Hepatitis B is an anthroponotic infection, predominantly with a parenteral mechanism of infection, which can occur in the form of viral carriage, acute and chronic forms and is characterized by liver damage with the possible development of acute liver failure, chronic hepatitis, liver cirrhosis and primary liver cancer (hepatocellular carcinoma).

Infection of the human body occurs through direct penetration of HBV into the blood (during any invasive interventions and transfusion of blood and its preparations), or through the mucous membranes and skin during childbirth, during sexual and close household contacts. Once in the liver, HBV penetrates the hepatocyte, where it begins to multiply intensively.

The development of the infectious process can occur in two ways: replicative and integrative. The replicative form of infection leads to the development of acute or chronic hepatitis and liver cirrhosis, while the integrative form leads to the development of “healthy” virus carriage, inactive chronic hepatitis, liver cirrhosis and primary hepatocarcinoma.

Serological diagnosis and prognosis of HBV infection is based on the identification of virus antigens and antibodies to it. But, as it turned out, in the hepatitis B virus population there are enough mutant strains (HBeAg-negative) that are not detected by conventional serological tests. Therefore, the PCR method for diagnosing HBV infection is extremely important.

Why is it important to make a qualitative determination of hepatitis B virus DNA?

PCR allows you to determine the DNA of the hepatitis B virus in the test material both qualitatively and quantitatively. Qualitative determination of the hepatitis B virus in material (whole blood, serum, plasma, liver biopsy) confirms the presence of the virus in the patient’s body and thereby establishes the pathogenesis of the disease. A quantitative method for determining the content of viral DNA in plasma provides important information about the intensity of disease development, the effectiveness of treatment, and the development of resistance to antiviral drugs. HBV PCR is clearly necessary to judge viral replication. Viral DNA in blood serum is detected in 50% of patients in the absence of HBeAg. Detection of HBV DNA in the blood is of great importance for the prognosis of acute HBV. It has been established that persistence of HBV DNA for more than 8 weeks after the onset of the disease indicates a chronic process, while elimination of viral DNA during the first 2 weeks of the disease correlates with complete recovery.

Qualitative determination of hepatitis B virus DNA by PCR.

Analytical indicators: detection of hepatitis B virus DNA by polymerase chain reaction (PCR) in blood plasma.

The determined fragment is a unique DNA sequence of the gene for the structural protein of the hepatitis B virus. The specificity of the determination is 98%. The detection sensitivity is at least 80 viral particles in 5 μl of a processed (DNA extraction) sample.

Thus, the determination of HBV DNA in a material is the most important analysis, which, together with other laboratory tests, allows one to diagnose an infection, determine the nature of the infectious process, and act as a criterion in carrying out therapy and assessing its effectiveness.

For what diseases is DNA of the hepatitis B virus done, qualitative determination?

Indications for the purpose of analysis:

Preventive screening studies (hepatitis B virus DNA is the earliest marker in acute infection).

Studies in HBsAg-negative infection.

Surveys of contact persons.

Diagnosis of hepatitis of mixed etiology - identification of the leading virus.

Detection of the stage of active viral replication in a chronic condition.

Monitoring of therapy.

Cirrhosis of the liver.

Weakness, malaise, fatigue, loss of appetite, nausea, heaviness in the right hypochondrium, enlarged liver, as well as pain in muscles and joints.

Quantitative determination of hepatitis B virus DNA by PCR on a Cobas TaqMan automatic analyzer.

Detection of hepatitis B virus (HBV) DNA using polymerase chain reaction (PCR) and determination of viral load in blood serum.

the test confidence range is now 54.5 – 1.10 E8 IU/ml;

Quantitative characterization of HBV DNA content in clinical samples is important for assessing the effectiveness of antiviral therapy.

Preparation

No special preparation is required for the study.

Indications

Positive qualitative test for the presence of hepatitis B virus DNA in blood serum.
Determination of patient treatment tactics.
Accurate assessment of treatment effectiveness.

Interpretation of results

low viremia - up to 10 4 copies/ml;
high - over 10 6 copies/ml.

Units of measurement (on the Cobas TaqMan auto analyzer): IU/ml - the amount of detected hepatitis B virus DNA.

Reference values: normal - “not detected”.

All about quantitative testing for hepatitis B

Hepatitis B is a viral infection that affects the liver. Today, about 300 million people around the world are carriers of hepatitis B.

In some, the virus progresses to cirrhosis of the liver or hepatocellular carcinoma (the first stage of cancer). New antiviral disease research strategies have two objectives:

determine how sensitive the body is to the viral load;
determine how resistant the virus is to medications and other medical interventions.

Features of tests for hepatitis

Serological tests are aimed at identifying antigens and antibodies in blood serum, but this method is unreliable. Therefore, scientists developed the polymerase chain reaction (PCR) method. It allows you not only to qualitatively determine the presence of the virus (is it present or not), but also quantitatively (how many antibodies are present in the blood serum).

Before collecting blood, the doctor should find out:

whether the patient took drugs intravenously;
whether the person was a donor or a recipient (to whom the blood was infused);
whether the patient underwent surgical interventions;
whether the patient’s skin was damaged;
whether the person is confident in the health of his sexual partner;
Have you had contact with someone infected with the hepatitis B virus?

In acute hepatitis, a positive result can be detected 1-2 weeks after incubation by PCR.

Blood tests to detect hepatitis B virus (HBV) are taken when:

Diagnosis of acute hepatitis in:
- incubation period (1-2 weeks);
- intensive period of illness (3-4 weeks);
- period of early recovery.
Diagnosis of CHV (chronic viral hepatitis).
For mixed hepatitis.

Tests are also regularly taken from people at risk, these include:

people who need frequent blood transfusions;
patients undergoing ongoing blood purification procedures for renal failure;
people with AIDS or HIV infection;
pregnant women;
healthcare workers exposed to blood;
patients with symptoms of liver disease.
those being treated for cirrhosis, cancer and other liver diseases.

Preparation for quantitative analysis requires compliance with the following rules:

do not smoke 1 hour before the test;
do not eat 4 hours before the procedure;
It is best to take a quantitative analysis after passing a qualitative one;
do not drink alcohol the day before the test.

Effective therapy for the disease affects the reduction of the amount of viral DNA in the serum. Six months after the start of treatment, the amount of virus should decrease by 2-3 orders of magnitude. If the test results have not changed over time, or, on the contrary, have become worse, then the entire therapy must be changed, and acute hepatitis is automatically renamed chronic.

When hepatitis virus DNA enters the body, infection can occur in 2 ways:

replicative (acute or chronic hepatitis develops, which in the future, without medical intervention, leads to liver cirrhosis);
integrative (development of asymptomatic, inactive virus carriage, which will still lead to cirrhosis or hepatocellular carcinoma).

How is the blood collection procedure performed? The doctor tightens the patient’s forearm with a tourniquet and inserts a needle into a vein on the elbow, then draws the blood into a syringe and pours it into a special tube. The results will be ready in a few days, the period depends on the place where the patient is tested.

The material is blood serum, lymphocytes, hepatobiopsy, which is placed in a test tube with a well-screwed cap. But the result can be affected by contaminated material for sampling, overexposure of the material (it is stored for up to 24 hours at a temperature of no more than +4 ° C).

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Quantitative analysis methods

There are several methods of quantitative analysis, including PCR, ELISA, and biochemistry.

PCR (polymerase chain reaction).

This is a test for the surface antigen protein that makes up part of the outer shell of the virus. After viral particles enter the body, they begin intensive reproduction on the surface of the liver, destroying healthy cells of the organ. New molecules are released into the blood.

Based on this, the level of antibody concentration in the serum is examined and it is determined whether there is infection with hepatitis B or not.

The material for the study is taken on an empty stomach. PCR is carried out in a machine called a cycler.

PCR analysis algorithm:

First, DNA is split into 2 independent chains by increasing the temperature of the material for research in a thermal cycler.
The material is cooled slightly with nitrogen. Primers find the desired sequences in the DNA molecule and attach to them.
Elongation is the third step where two primers are added to two strands of DNA. With the help of the enzyme DNA polymerase, the DNA section is completed from the primer that was previously attached to it. Thus, at the site in the primer region in each of the DNA strands, a second complementary DNA strand is formed.

Subsequently, these steps are repeated many times, and in a few hours cycles are completed, billions of copies of the desired sample are formed. If there are such copies, then their number is calculated per 1 ml of material for analysis.

In addition to PCR, there is an ELISA (enzyme-linked immunosorbent assay) method. It determines not only surface antibodies, but also those located inside and interrelated with previous antigens, as well as their quantity.

Biochemistry

Another method of quantitative analysis. When a virus enters the body from damaged liver cells, enzymes are released; if their amount is higher than normal, then we can talk about infection. In addition, you need to determine the viral load (DNA-HBV), take liver tests (for bilirubin with fractions, ALT, AST, alkaline phosphatase, gamma-GT). It is mandatory to visit an infectious disease specialist, who, if necessary, refers the patient to fibroelastometry and selects a course of treatment.

Biochemistry - quantitative analysis

Real-time PCR

In this method, the search for copies occurs after each cycle, and not after 35-45. The method works in the same way as PCR, it allows you to determine the number of copies in a sample for research. Thus, the analysis time is noticeably reduced, while a 100% result is guaranteed.

Hepatitis B DNA detection

Establishing the amount of hepatitis B DNA is very important, since with a low level the prognosis of the disease is more favorable than with a higher level. HBV DNA concentration is measured in copies/ml or me/ml

1 me/ml = 2.83×10 copies/ml

The results of this analysis may be as follows:

Virus DNA is not detected if it is not in the body or the concentration is very low, 90 me/ml = 200 copies/ml;
Viral DNA was detected in quantities below the normal concentration, 2×106 copies/ml;
Viral DNA is in average quantity, at 2×106-2×109 copies/ml;
The viral DNA is more than 2x109 copies/ml, the concentration of antibodies is high, which indicates the progress of the disease.

If the test result is positive, then the following is diagnosed:

If the result is negative, then:

In rare cases, a negative DNA test result for the hepatitis B virus indicates a rapid and malignant course of the disease.

The decision to get tested for the virus is made by:

Quantitative analysis is mandatory when diagnosing the disease and should take place before starting therapy. But it must be remembered that hepatitis B is sometimes closely intertwined with hepatitis D, which can create additional difficulties when conducting such an analysis. Therefore, the following complications should be taken into account:

coinfection – infection occurs with two types of hepatitis B and D at once. This disease is much more complex and painful. This disease does not have a chronic form, because in most cases the disease ends in death;
superinfection - acute HBV joins previously dormant hepatitis D, in this case the disease becomes chronic with an unfavorable prognosis.

PCR for hepatitis B

PCR diagnostics allows not only to determine the presence of hepatitis B virus in the blood and its etiology, but also to evaluate its activity. Detection of the viral load is especially important for selecting effective treatment; if it is too high, the likelihood of recovery decreases. What is the essence of the polymerase chain reaction method?

The essence of PCR diagnostics

DNA virus in the blood can be detected by PCR for hepatitis at the end of the incubation period, at this time, against the background of increased levels of transaminases, HBsAg can be detected, after which HBeAg appears.

With a qualitative determination, you can accurately diagnose whether there is hepatitis or not. Normally, there should be no viral DNA in the blood.
The quantitative method allows you to determine the intensity of the disease and the reproduction of the virus.

The analysis has very high sensitivity and reliability. Venous blood is taken as biological material. Thanks to modern technologies, PCR can detect the virus at a concentration of 5x copies/ml in the blood. Based on the results of the analysis, knowing the norm, you can judge the viral load and make a prognosis for treatment.

Important! It is the DNA of the virus that contributes to the development of cirrhosis and other chronic liver diseases.

DNA detection using PCR is necessary in the following cases:

Doubts when setting up the final analysis after taking the tests.
Determination of the acute stage of the disease.
Detection of latent forms of hepatitis.
Evaluation of effectiveness after antiviral therapy.

How to decipher the results obtained after PCR diagnostics?

PCR quantitative

By quantitative assessment, you can not only determine the presence of the virus, but also find out the viral load if the PCR shows a positive result.

A quantitative method is needed to find out the following information:

Quantitative assessment is very important when diagnosing chronic hepatitis. In this case, all indicators will not be within normal limits. The transaminase level will be increased, the virus activity index will be more than 4, and the virus DNA concentration will be more than 105 DNA copies/ml. If the virus concentration is lower and the transaminase level is normal, then we can talk about passive carriage.

Before prescribing antiviral treatment, undergoing PCR diagnostics, namely a quantitative examination, is a prerequisite to determine the viral load.

They do an analysis every 3 months, and if the viral load has increased 10 times, then we can talk about the virus’s resistance to treatment.

Quantitative analysis provides very important information, because you can determine how much DNA of the pathogen is contained in the blood. The greater the amount, the greater the viral load and the more severe the patient’s condition.
If the viral load decreases after treatment, one can judge its effectiveness.
In some cases, PCR for hepatitis is an indication for additional examination, such as a liver biopsy. If ALT levels are elevated, PCR is performed. The interpretation of the tests is as follows: if the viral load is more than 10 5 copies of DNA/ml, and the ALT level exceeds the norm, but no more than 2 times over the course of six months, then such a patient needs a biopsy. For severe inflammation or fibrosis, antiviral treatment is indicated. If the ALT level exceeds the norm by more than 2 times with a high viral load, then treatment is immediately prescribed without additional diagnostics.

Only good specialists will be able to correctly interpret the results of quantitative PCR.

PCR qualitative

A qualitative analysis allows you to determine the presence of hepatitis virus in the blood. Normally it should be absent. Thanks to this method, you can accurately establish the diagnosis.

Qualitative PCR analysis gives a 100% accurate result.

No other method provides such reliable data. In more than half of the cases, after diagnosis, viral DNA is detected in the absence of antigen. It is very important to establish a diagnosis at the initial stage of the disease.

It has been proven that if the DNA of the hepatitis B virus actively multiplies in the human body for two months, the disease becomes chronic. If you start fighting the virus within the first weeks after infection, then the chances of a full recovery are very high.

How is the survey results deciphered using the qualitative method?

Decoding the analysis is very simple:

Normally, the result should be negative, that is, no viral DNA was detected.
A positive result indicates the presence of hepatitis.

The analysis helps to identify the virus, determine its genotype and begin timely treatment. Very often, in the presence of DNA, along with a qualitative assessment, a quantitative assessment is also carried out.

How to prepare for the examination?

When testing for hepatitis using PCR diagnostics, blood is taken from a vein. It is best to undergo the examination in the morning, since blood must always be taken on an empty stomach (at least 8 hours must pass after the last meal).

In order for the analysis to be as reliable as possible, the patient must be aware of some factors that may affect the final result:

Before donating blood from a vein, you need to rest for 20 minutes.
In addition to the fact that the test is taken on an empty stomach, for another 12 hours you should not drink alcohol, smoke, play sports or eat fatty foods.
Some medications that the patient is forced to take can affect the final result. The laboratory technician must be aware of the drug treatment case.
If blood needs to be donated to a child under 5 years of age, then he should be given water every 10 minutes half an hour before diagnosis. a glass of boiled water.

It is very important to undergo PCR in a clinic that has proven itself well. Indeed, despite the fact that PCR determines the presence of the virus with an accuracy of 100%, this figure can drop to 95% if we take into account the human factor.

Unqualified laboratory technicians or a bad reputation of the clinic can cause false results.

Hepatitis B virus DNA, quantification

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What is hepatitis B virus DNA quantification?

HBV is a complex DNA virus and belongs to the Hepadnaviridae family, genus Orthohepadnavirus.

Quantitative determination of hepatitis B virus DNA by PCR.

Analytical indicators: detection of hepatitis B virus DNA using polymerase chain reaction (PCR) and determination of viral load in blood plasma.

The fragment being determined is a unique DNA sequence of the gene for the structural protein of the hepatitis B virus. The specificity of the determination is 98%. The sensitivity of detection is from 2x105 IU/ml to 5x103 copies/ml of viral particles in blood serum.

Quantitative characteristics of hepatitis virus DNA content are important for assessing the effectiveness of antiviral therapy and have prognostic significance for determining the chronicity of hepatitis B: with an initially low level of viremia (HBV DNA less than 2x105 copies/ml (2x105 IU/ml)), the percentage of acute hepatitis B chronicity is close to zero . When the HBV DNA concentration is from 2x105 copies/ml (2x105 IU/ml) to 2x10° copies/ml (8x105 IU/ml), chronicity of the process is observed in 25-30% of patients, and with a high level of viremia in the patient (more than 2x10° copies/ ml (8x105 IU/ml)) acute hepatitis B most often turns into chronic. An 85% decrease in hepatitis B virus DNA concentration by the third day from the start of therapy is a rapid and accurate parameter for predicting the effectiveness of therapy.

Types of PCR for hepatitis B and interpretation of results

Polymerase chain reaction is a type of blood test that allows you to accurately detect the hepatitis B virus. A special feature of the method is that it is able to show HBV in the early stages - within a month after infection, while a specific antigen in a patient is detected only after 2 months . PCR can detect not only a regular DNA virus, but also mutant strains that no other analysis can detect.

When deciphering PCR for hepatitis B, the results are compared with standard values. Based on this, conclusions are drawn about the presence of the disease (positive result), the concentration of the virus (activity) or the absence of the disease (negative result). Data processing takes 2 weeks.

Types of PCR for viral hepatitis B

There are 2 types of diagnostics that are prescribed for hepatitis B. Qualitative PCR makes it possible to say with 100% accuracy whether HBV is in the blood or not. Quantitative PCR shows the activity of the virus, that is, how many HBV DNA fragments are contained in 1 ml of blood.

Qualitative PCR and its interpretation

A blood test for hepatitis B using the PCR method can only reveal the presence of DNA fragments. It is prescribed when the patient suspects infection with HBV or when other diagnostic methods, such as serological tests, do not provide a clear answer. Decoding the results when conducting a qualitative analysis is very simple:

negative result - there is no virus;
positive result - the virus has been detected.

The purpose of high-quality PCR is to confirm or refute the presence of the hepatitis B virus, as well as to conduct early diagnosis. If HBV is detected within 1-2 weeks after infection, then treatment will be most effective. With the active development of HBV for 2 months or more, the disease becomes chronic and cannot be completely cured.

In the case when qualitative PCR for hepatitis shows the presence of the virus, the next step is quantitative diagnosis.

Quantitative PCR

To prescribe effective treatment or evaluate the results of therapy, simply knowing that the patient has HBV is not enough. It is necessary to quantify the DNA value of the hepatitis B virus in 1 ml of blood, that is, the viral load on the body. This type of PCR for hepatitis B is carried out in the following cases:

Before prescribing treatment.
To assess the effectiveness of therapy.
To determine the development of virus resistance to drugs taken.
To determine the stage at which the disease is located.

The unit of measurement for the virus is international units (IU/ml) or the number of DNA copies/ml, that is, the number of DNA fragments per 1 ml of blood. The ratio between the two units depends on the test system chosen and ranges from 2 to 7 copies/ml in 1 IU/ml. If the system is unknown, then the average value is used for recalculation:

1 IU/ml = 5 copies/ml

If a quantitative analysis is immediately done, then the limit norm for diagnosing the disease is 75 IU/ml. If the result is higher than this indicator, a diagnosis of HBV is made; if the result is lower, HBV is not detected.

Interpretation of quantitative PCR

For hepatitis B, PCR standards are the following values of viral load (viremia), copies/ml:

< 10^3 - нагрузка очень низкая, вирус ведет себя неактивно;
from 10^4 to 10^5 - the load is low, the virus is usually inactive;
10^6 - the load is moderate, the virus is active;
10^7 - high viremia, the virus is highly active.

Knowing quantitatively how the virus behaves in the body, you can determine the form of the disease (chronic, acute or carriage), the need for additional research, the reaction of the body and the virus to treatment:

Chronic hepatitis B is diagnosed if the transaminase value is higher than normal, the virus activity is more than 4, and the concentration of DNA copies per milliliter of blood is 10^5 copies.
Carriage is determined by the following picture that the results give - concentration< 10^5 копий/мл, а уровень трансаминазы равен стандартному значению.
HBV resistance is constantly monitored during treatment. Tests are taken every 3 months. If at one point the viremia during hepatitis increases 10 times, this means that HBV has adapted to the drugs used - it has become resistant.
A favorable course of treatment will show a decrease in viremia by 85% 3 days after the start of therapy.
The patient receives a referral for a liver biopsy only when the ALT (alanine aminotransferase) level exceeds the normal value no more than 2 times in 6 months, and the PCR for hepatitis B shows viremia > 10^5 copies/ml. When, with high viral activity, an increase in ALT levels is observed more than 2 times in six months, then antiviral therapy is immediately prescribed.

To find out how DNA hepatitis will behave in terms of chronicity, that is, the transition to a chronic form from an acute form, the results of PCR diagnostics are also used:

HBV DNA< 2 х 10^4 копий/мл означает, что острая стадия не перейдет в хроническую форму;
HBV DNA ranges from 2 x 10^4 to 2 x 10^6 copies/ml, which means that every 4th patient will have chronic HBV, that is, chronicity is 25-30%;
HBV DNA > 2 x 10^6 copies/ml means that the acquisition of a chronic form of the disease is inevitable.

Preparing for the examination

To determine the presence of HBV using PCR, a blood test from a vein is performed. The procedure is always carried out strictly on an empty stomach and it is best to get to the laboratory in the morning, when the natural rest time from food is 8 hours. In order for the examination results to be correct, you need to know in advance how to prepare for going to the clinic correctly. The additional steps are:

Avoid fatty foods, smoking, alcohol and sports 12 hours before the procedure.
Rest for 20 minutes immediately before donating blood.
Before starting the procedure, you must inform the nurse about the medications you are taking.

Attention! If the diagnosis is carried out on a child under 5 years old, then half an hour before the procedure he should be given 1 glass of boiled water every 10 minutes.

Summing up

Today, PCR diagnostics for detecting hepatitis B is the most accurate method. The method allows you to see hidden and mutated forms of the disease, identify the disease in the first weeks after infection and, accordingly, carry out effective treatment. Depending on the situation, qualitative or quantitative PCR is carried out, which are aimed at identifying HBV and establishing its activity. To undergo an examination, you need to contact a virologist, infectious disease specialist or hepatologist.

HBV, DNA quantitative [real-time PCR]

A test to identify the causative agent of hepatitis B (HBV), during which real-time polymerase chain reaction (RT-PCR) is used to determine the presence of genetic material (DNA) of the virus and its amount (viral load) in a blood sample.

HBV DNA may be detected at concentrations below the lower limit of the linear concentration range. The linear concentration range is the range within which the copy number of a pathogen can be accurately counted. For this assay, the linear range of HBV DNA concentrations determined by the detection cycle is 7.5 x 10 2 – 1.0 x 10 8 copies/ml of sample.

Hepatitis B virus (HBV), DNA quantification.

Hepatitis B Virus DNA, Quantitative, Real-Time PCR, Blood; HBV Viral Load; Hepatitis B Virus DNA Quant.

Real-time polymerase chain reaction.

Copy/ml (copy per milliliter).

What biomaterial can be used for research?

How to properly prepare for research?

Do not smoke for 30 minutes before donating blood.

General information about the study

Hepatitis B virus (HBV) is an infectious liver disease caused by the DNA-containing hepatitis B virus (HBV). Among all the causes of acute hepatitis and chronic viral infection, the hepatitis B virus is considered one of the most common in the world. The actual number of people infected is unknown, since many people have an infection without specific symptoms and do not seek medical help. The virus is often detected during preventive laboratory tests. According to rough estimates, about 350 million people in the world are affected by the hepatitis B virus and 620 thousand die from its consequences every year. In Russia, the number of HBV carriers exceeds 5 million people.

The source of infection is a patient with HBV or an asymptomatic virus carrier. HBV is transmitted through blood and other body fluids. You can become infected through unprotected sexual contact, the use of unsterile syringes, blood transfusions and organ transplants; a child can be infected by the mother during or after childbirth (through cracks in the nipples). The risk group includes: medical workers who may have contact with the patient’s blood, patients receiving hemodialysis, injection drug users, people who are promiscuous, children born to mothers with HBV.

The incubation period of the disease is from 4 weeks to 6 months. Viral hepatitis B can occur either in mild forms, lasting several weeks, or as a long-term chronic infection. The main signs of hepatitis: jaundice of the skin, fever, nausea, fatigue, in laboratory tests - abnormal liver function and specific antigens of the hepatitis B virus. An acute disease can quickly lead to death, develop into a chronic infection or end in complete recovery. It is believed that after suffering from HBV, stable immunity is formed. Chronic viral hepatitis B is associated with the development of cirrhosis and liver cancer.

There are several specific tests to detect existing or past viral hepatitis B. To confirm the presence of infection and clarify the period of the disease, they use the determination of virus antigens, antibodies to them and viral DNA.

The polymerase chain reaction is highly sensitive and specific. The PCR method can determine the DNA of the virus qualitatively or quantitatively. Thanks to the qualitative method, the presence of the hepatitis B virus in the body and its active reproduction are confirmed. Quantitative determination of the viral load allows us to assess the intensity of the disease, the effectiveness of the therapy, or the development of resistance to antiviral drugs.

There is a relationship between the concentration of the virus in the blood and the outcome of acute viral hepatitis B. With a low level of viremia, the likelihood of the infection becoming chronic is close to zero, and the infected person is not dangerous to others. With a high viral load (5 copies/ml), chronicity occurs frequently and the patient is a potential source of infection. A connection has been proven between the amount of viral DNA in the blood serum, the presence of HBeAg, increased ALT and the development of liver cirrhosis and hepatocellular carcinoma (liver cancer).

The effectiveness of antiviral therapy is assessed by reducing the amount of viral DNA in the blood. 3-6 months after the start of treatment, the viral load with an adequate therapeutic response should decrease by 1-2 orders of magnitude. The absence of a decrease in the amount of virus or its increase during treatment requires a review and change in therapy.

Quantitative determination of hepatitis B virus DNA, together with the clinical picture of the disease and biochemical parameters, markers of infection, as well as the result of liver puncture biopsy, allows us to predict the disease and assess the need for antiviral therapy.

What is the research used for?

To predict the course of viral hepatitis B.
To confirm the chronic form of viral hepatitis B.
To identify carriers of viral hepatitis B and monitor virus replication activity.
To identify hidden and mutant strains of the hepatitis B virus.
To assess the effectiveness of antiviral therapy for hepatitis B and make decisions on further treatment tactics.

When is the study scheduled?

With qualitative detection of hepatitis B virus DNA.
For acute and chronic viral hepatitis B.
For mixed hepatitis.
Before and during antiviral therapy.

“Not detected” – hepatitis B virus DNA was not detected or the value is below the sensitivity limit of the method (200 copies/ml = 120 IU/ml).
10^5 copies/ml (6*10^4 IU/ml) – high viral load, active infectious process (antiviral therapy is required).
> 1.0*10^8 copies/ml (>6*10^7 IU/ml) – hepatitis B virus DNA was detected in concentrations above the linear concentration range.

The linear range of hepatitis B virus DNA concentrations determined by the detection amplifier is 7.5*10^2 – 1.0*10^8 copies/ml (4.5*10^2 – 6*10^7 IU/ml).

What can influence the result?

Quantitative determination of hepatitis B virus DNA is a mandatory test before prescribing antiviral therapy. During the course of treatment, the analysis must be repeated after 3-6 months.
Viral hepatitis B is often combined with viral hepatitis D.

Who orders the study?

DNA and PCR studies of the hepatitis B virus are the most sensitive tests that can detect molecular traces of DNA in liver tissue that identify the presence of the virus. The essence of the method is to repeatedly copy a DNA fragment belonging to the causative agent of the disease.

Hepatitis B type PCR is a polymerase chain reaction of an external chemical reagent with the biological material being studied, aimed at identifying viral DNA molecules in the blood and their quantitative indicators per unit volume. Analysis is prescribed in the following cases:

if signs of hepatitis B occur in the patient;
during diagnostic procedures for mixed hepatitis;
to clarify the effectiveness of therapy.

For PCR analysis, venous blood is taken. Before the study, you must abstain from eating for approximately 8 hours.

The normal result when detecting DNA from a virus is considered to be “not detected.” It means that there is no virus in the blood or its content is insignificant. The final diagnosis can only be made by a specialist based on a comprehensive biochemical and immunological analysis of blood plasma for hepatitis type B. The doctor prescribes a course of therapy. Self-medication entails too much risk.

The PCR method makes it possible to detect pathogen DNA even if its presence is minimal in the selected blood sample. In addition, the analysis allows you to detect antigens that have already been produced by the virus. Each of the antigens indicates a certain stage of the disease:

HBsAg - the virus is present at the moment.
Anti-HBcor is not currently present, but was present in the past.
Anti-HBs - there are antibodies to hepatitis B in the blood (optionally - herpes, HIV, chlamydia, mycoplasmosis, candidiasis, tuberculosis, tick-borne encephalitis and many other diseases).

Pathology and its signs

Hepatitis B or HBV is a viral infection that affects liver cells. It is dangerous not only because of its direct effect on the organ parenchyma, which weakens the barrier function and general immunity. More severe consequences are also possible, such as liver cirrhosis and the occurrence of malignant neoplasms.

HBV is transmitted through the blood of a person infected with hepatitis, his excrement, through contact with mucous membranes, especially during sexual intercourse, skin lesions, through unsterile syringes during blood transfusions or during abdominal operations. It is possible for a newborn to become infected by his sick mother by touching the cracks in her nipples.

The following symptoms are typical for hepatitis B:

increasing fatigue with light loads;
unusual fatigue;
heaviness in the right hypochondrium;
decreased appetite, nausea;
yellowness of the skin, sclera of the eyes;
light stool;
dark urine;
painful sensations in the joints.

The incubation period after which these signs begin to appear ranges from a month to six months. This is the period of accumulation of virus cells in the hepatic bloodstream. When their concentration reaches a critical mass, the virus breaks through the membranes of hepatocytes and continues to multiply inside the cells. As a result, liver cells die, cirrhosis develops, which, in the absence of proper therapeutic measures, leads the patient to death. A timely blood test for PCR and hepatitis B DNA allows you to avoid negative ones.

Indications for PCR are:

a positive DNA test confirming the diagnosis of hepatitis B;
acute and chronic hepatitis;
mixed hepatitis;
quantitative determination of virus in blood serum;
development of effective therapy tactics.

Preparing for analysis

Preparation for diagnosis using PCR is determined by the type of biomaterial. So blood and urine are donated only in the first half of the day, on an empty stomach. Smears and scrapings are also taken from the urogenital area. The only restriction for women is menstruation. For both sexes, sexual intercourse on the eve of the study is limited.

complete peace of mind, absence of any worries. To ensure this, the test can be taken even at home. You just need to call a doctor with the appropriate equipment;
abstaining from food, alcohol, smoking, physical activity at least 8-12 hours before the analysis, otherwise the study data will be distorted;
15-20 minutes of complete relaxation before blood sampling;
stopping medication (if this is not possible, the laboratory should be notified);
Children under the age of five should be given boiled water to drink in small portions for half an hour before the test.

How the research works

PCR analysis is based on the principles used in molecular biology. They involve the use of special enzymes that cleave and copy sections of DNA and RNA of the infectious agent found in the blood. After this operation, the obtained samples are identified according to the database available in the laboratory. At the same time, the type of infection and the concentration of the pathogen in the blood are detected.

To conduct the research, a thermostat is used, a thermostat capable of heating and cooling tubes with biomaterial. This is necessary to activate replication processes. The reliability of the analysis depends on the accuracy of temperature parameters.

Decoding the results

A positive result for a patient is a laboratory result of “not detected.” It means the absence of hepatitis B DNA in the blood. In digital terms, this is approximately 120 IU/ml, or 200 copies per same unit of blood volume. This value is the norm. The requirement for biological material is that it must not be contaminated.

Decoding other analysis results allows us to make the following diagnoses:

The concentration of hepatitis B DNA in the blood is more than 200 copies, but below the critical level. There is no obvious pathology.
Moderate viremia, reaching up to 500 copies per ml. The choice of treatment depends on the results of the liver biopsy and the degree of deterioration in liver function.
Increased concentration of the virus (more than 500 copies/ml). The inflammatory process is in the active phase and requires intensive antiviral therapy.
Limit viral load (over 800 copies per ml.). Therapeutic measures are developed based on the patient’s condition and are entirely aimed at improving it as much as possible.

Determination of quantitative indicators of hepatitis B DNA content during PCR is the basis for choosing a treatment course. After three to six months, tests are done again to check the effectiveness of antiviral therapy.

Feature of polymerase chain reaction

The PCR technique uses two approaches to determine the DNA belonging to the virus: qualitative and quantitative.

A qualitative approach confirms or denies the presence of hepatitis B. PCR is prescribed at the request of a patient who suspects that he is infected with HBV infection or in cases where early diagnosis by other means does not give clear results.

If the hepatitis B virus can be detected no later than two weeks after it enters the body, a positive prognosis for treatment is most likely. In the future, the disease will begin to progress. After about two months it will enter the chronic phase. A complete cure in this case will become impossible.

The quantitative PCR approach is used only when a virus is detected. He evaluates:

specific viral load on the liver;
intensity of disease progression;
resistance to antiviral drugs;
effectiveness of treatment.

These indicators have direct practical significance. Especially quantitative DNA indicator. The prognosis in the acute phase of hepatitis B depends on the concentration of the virus in the liver bloodstream. Thus, a low concentration of viral cells almost completely eliminates the transition of acute to chronic hepatitis. Such patients do not pose a danger to others. A high viral load contributes to the chronicity of the pathological process and the possibility of hepatitis infection of people in contact with the patient.

There is also a connection between the concentration of virus genes in the blood and cirrhotic degeneration of the liver and the occurrence of cancer of its parenchyma.

The effectiveness of therapy with antiviral drugs is determined by the degree of reduction in the concentration of hepatitis B DNA in the hepatic bloodstream. A sufficient result is considered to be a reduction in viral load by 1-2 times with intensive treatment for three to six months. Lower rates require a change in the treatment course.

The prognosis for the development of pathology and the prescription of appropriate therapy is based on:

quantitative volume of liver virus DNA in blood serum;
accompanying biochemical indicators (for example, the level of leukocytes in the blood and the content of bilirubin there);
markers of viral infection;
liver biopsy results.

General purposes of PCR

Polymerase chain reaction tests help the doctor:

detect virus carriers;
predict the development of hepatitis type B;
make sure that the disease becomes chronic;
monitor the activity of the virus at all phases of development;
identify mutant and latent strains of hepatitis B;
evaluate the effectiveness of treating the virus when choosing a certain tactic and change it if it is not successful.

Advantages of the method

Unlike other tests that are used to diagnose hepatitis B, PCR has:

Exceptional sensitivity. The technique allows the researcher to detect the causative agent of the disease, even if the concentration of its DNA molecules in the blood is minimal. This allows you to take preventive and therapeutic measures at an early stage of the disease, when outwardly it is not yet expressed by any symptoms.
Wide range of possibilities. PCR can detect not only the causative agent of viral hepatitis B, but also a number of other pathologies.
Universality of approaches. The biomaterial for research can be blood, saliva, cells of mucous membranes and skin.
Efficiency. The conclusion will be ready exactly one day after the blood is taken (the test result itself is received no later than 7 hours later).
Credibility. If all conditions are met, the PCR method never shows false data.
Affordability of research. The cost of PCR is only slightly higher than that of conventional tests that use blood as a biomaterial.

The only cost of the research is its technological complexity. To perform high-quality PCR, only pure biomaterial, modern equipment and well-trained personnel are required. It is preferable to seek diagnostics using this method in specialized laboratories.

Viral hepatitis B is a disease for which complete recovery is possible only in the earliest stages of development. Ultra-sensitive diagnostics using the PCR method helps to identify the disease in this phase. The first symptoms that raise suspicion of liver damage by a virus should be a reason to contact a specialist. If the diagnosis is confirmed, treatment should begin immediately. This will improve your prognosis and increase your likelihood of maintaining a healthy liver for years to come.

Synonyms: HBV DNA

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Description
Decoding
Why Lab4U?

Period of execution

The analysis will be ready within 7 days, excluding Saturday and Sunday (except for the day of taking the biomaterial). You will receive the results by email. mail immediately when ready.

Completion time: 2 days, excluding Saturday and Sunday (except for the day of taking biomaterial)

Preparing for analysis

In advance

Do not take a blood test immediately after radiography, fluorography, ultrasound, or physical procedures.

Discuss with your doctor the intake of medications the day before and on the day of the blood test, as well as other additional preparation conditions.

The day before

24 hours before blood collection:

Limit fatty and fried foods, do not drink alcohol.

Avoid heavy physical activity.

For at least 4 hours before donating blood, do not eat food, drink only clean, still water.

On the day of delivery

Do not smoke 60 minutes before blood collection.

Be in a calm state for 15-30 minutes before taking blood.

Analysis Information

Index

Hepatitis B can be asymptomatic for a long time, becoming chronic. Chronic viral hepatitis B can lead to cirrhosis and liver cancer. While a person does not know that he is sick, it is dangerous for him and others.

First of all, after infection, hepatitis B can be detected by a blood test for the DNA of the hepatitis B virus. It will show infection after 10-12 days. This test will help check the possible infection of a person at risk.

For the purpose of prevention, a test for hepatitis B can be taken as a complex.

Appointments

A quantitative method for determining the DNA content of the hepatitis B virus in the blood makes it possible to assess the intensity of the disease, the effectiveness of treatment and the development of resistance to antiviral drugs.

Specialist

Prescribed by a therapist, endocrinologist, infectious disease specialist, hepatologist.

Research method - Real time PCR

Material for research - Venous blood with EDTA

Composition and results

Hepatitis B virus, DNA determination (quantitative)

Viral hepatitis B (HBV - hepatit b virus) is an infectious liver disease caused by the hepatitis B virus.

The most common type of hepatitis on the planet. In Russia, about 5 million people suffer from it, and a considerable number of people do not suspect that they are sick. Hepatitis B can be asymptomatic for a long time, becoming chronic. Therefore, while a person does not know that he is sick, it is dangerous both for his health and for the health of others. For this reason, it is recommended to take a test to detect hepatitis B regularly, once every six months. You can protect yourself from hepatitis B infection through vaccination. Vaccination provides protection against hepatitis B for 5 to 7 years. Before vaccination, after vaccination and 5 years after vaccination, it is necessary to examine the level of antibodies using an analysis -.

Hepatitis B is transmitted through biological fluids: semen, blood, plasma. If there was a probable case of infection (unprotected sexual intercourse, repeated use of a syringe, contact with infected blood, etc.), you need to take a test for hepatitis B in a month for a more reliable result.

The incubation period is from 4 weeks to 6 months. The incubation period is the length of time from the moment the virus enters the body until the symptoms of the disease appear. At the end of the incubation period, the levels of liver parameters (ALT, AST) increase, the liver and spleen enlarge, and the concentration of bilirubin increases to 2 - 2.5 times the norm, although this does not lead to darkening of the urine. It can occur both in the form of mild forms, lasting several weeks, and in the form of a chronic infection with a long-term course.

Symptoms of acute hepatitis: yellowness of the skin, fever, nausea, fatigue. Laboratory tests show signs of liver dysfunction and specific antigens of the hepatitis B virus. Acute hepatitis is the period of the first 3 months from the moment of infection. Acute hepatitis can occur with or without clear symptoms of hepatitis (jaundice, whitish stools).

An acute disease can occur quickly, be fatal, develop into a chronic infection, or end in complete recovery. It is believed that after hepatitis, stable immunity is formed. Chronic viral hepatitis B can lead to cirrhosis and liver cancer. In the process of the body's fight against hepatitis, autoimmune diseases can form: thyroiditis, chronic gastritis, Sjögren's syndrome, idiopathic thrombocytopenic purpura, periarteritis nodosa, glomerulonephritis, Guillain-Barre syndrome, rheumatoid arthritis, etc.

Diagnosis of acute hepatitis B

First of all, after infection, hepatitis B can be detected by a blood test for the DNA of the hepatitis B virus. During this period, the infected person is contagious, although symptoms of hepatitis may not be observed. However, this analysis may also be negative during a certain period (diagnostic or serological window) of the course of the disease. After some time (3-5 weeks) from the moment of infection, the body begins to produce

The diagnosis of hepatitis B is made based on the determination of several tests characteristic of various stages of the disease - the incubation period, the acute phase and convalescence.

A quantitative method for determining the DNA of the hepatitis B virus in the blood makes it possible to assess the intensity of the disease, the effectiveness of treatment and the development of resistance to antiviral drugs.

Determining the DNA of the virus is certainly necessary to establish the phase of virus development. Determination of viral DNA is very important for prognosis of the course of acute hepatitis B. It has been established that unchanged viral DNA in the blood for more than 8 weeks after the onset of the disease indicates a chronic process, while detection and initiation of treatment for viral DNA during the first 2 weeks of the disease correlates with complete recovery. Therefore, determining the DNA of the hepatitis B virus in the blood is the most important test, which, together with other laboratory tests, makes it possible to diagnose the infection, determine the nature of the infectious process, and assess the need for therapy and its effectiveness.

The sensitivity of the method is 255 DNA copies/ml or 150 IU/ml.

Interpretation of the results of the study "Hepatitis B virus, DNA determination (quantitative)"

Interpretation of test results is for informational purposes only, does not constitute a diagnosis and does not replace medical advice. Reference values may differ from those indicated depending on the equipment used, the actual values will be indicated on the results form.

Unit of measurement: DNA copies/ml, IU/ml

Reference values: <255 копий ДНК/мл; <150 МЕ/мл; 1МЕ = 1,7 копий ДНК/мл

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