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I. ДИАГНОСТИКА И ЛЕЧЕНИЕ
ON THE ISSUE OF LABORATORY DIAGNOSIS OF VIRAL HEPATITIS C
yflK 616.36-002-074
Abdilova G.B., Nurakhova A.D., Baichalova A.D., Abdigalieva G.K.
National Scientific Surgery Center under the name of A.N.Syzganov, Almaty, Kazakh Medical University Continuing Education, Almaty
ABOUT THE AUTHORS
Abdilova Gulnur Bekmurzaevna -
Head of the CDL,
e-mail address: gulnur_abdilova@mail.ru, telephone 87019911346
Nurakhova Alma Dandybaevna -
PhD, doctor-laboratory CDL, e-mail address: nad7788@mai.ru, telephone 87776850298
Abstract
In this paper there is the analysis of enzyme immunoassay research of viral hepatitis C markers in the data ex- -
tracted from the blood serum of patients treated and examined in National Scientific Surgery Center under the name of Keywords A.N.Syzganov in 2016. The article is devoted to the diagnosis of hepatitis C, as the viral hepatitis is a major public health linked immunosorbent assay, problem, affecting approximately 3.3% of the total population. viral hepatitis, viral hepatitis C.
С вирусты гепатиттщ зертханальщ диагностикасы мэселесше
Абдилова Г.Б., Нурахова А.Д., Байчалова А.Д., Абдигалиева Г.К.
«A.H.Cbi3faH0B атындаш ¥FXO» AK,, Алматы к,.,
Уздшз 6rniM беру бойынша Казак медицина университет!, Алматы к.
Ацдатпа
Жумыста 2016 жылы А.Н. Сызганов атындагы ¥лттьщ гылыми хирургия орталь^ы АКтексеруден еткен жэне емделген С гепатитi бар наукастардыц кан сарысуы иммундыкферментт1к маркерлер!нщ корытындылары бойынша сараптама щрпз'тщ. Макала Вирустык C гепатитiнiц диагностикасыныц сурактарына арналган, сондай-ак, вирустык гепатит жалпы алганда 3,3% бук'ш халыктыц денсаулыгына зиян келпрепн мацызды проблема болып табылады.
АВТОРЛАР ТУРАЛЫ
Абдилова Гупьнур Бекмурзаевна -
КДЗ мецгеруша,
Электронды адресг. gulnur_abdilova@mail. ru, телефон 8701991134
Нурахова Алма Дандыбаевна -
мг.к., КДЗ лаборант-дaрiгерi, электронды адресг. nad7788@mai.ru, телефон 87776850298
Туйш сездер
иммундыкферменттiк сараптама, вирусты гепатиттер, вирусты гепатит С.
К вопросу лабораторной диагностики вирусного гепатита С
Абдилова Г.Б., Нурахова А.Д., Байчалова А.Д., Абдигалиева Г.К.
AO «ННЦХ им. А.Н.Сызганова», г. Алматы,
Казахский медицинский университет непрерывного образования, г. Алматы
ОБ АВТОРАХ
Абдилова Гупьнур Бекмурзаевна -
Заведующая КДЁ,
электронный адрес: gulnur_abdilova@mail. ru, телефон 8701991134
Нурахова Алма Дандыбаевна - к.м.н., Врач-лаборант КДЁ, электронный адрес: nad7788@mai.ru, телефон 87776850298
Аннотация
В работе выполнен анализ данных иммуноферментных исследований маркеров вирусного гепатита C, выделенных из сыворотки крови пациентов, лечившихся и обследовавшихся в ННЦХ им. А.Н.Сызганова в 2016 г. Статья посвящена вопросу диагностики вирусного гепатита C, так как данный вирусный гепатит является важной проблемой общественного здравоохранения, затрагивая примерно 3,3 % всего населения.
Ключевые слова
иммуноферментный анализ, вирусные гепатиты, вирусный гепатит С.
To date, due to the intensification of the processes of globalization, diseases of infectious etiology, due to the high prevalence in the world, constitute a global threat. Including viral hepatitis C. According to WHO, about 150 million people in the world (approximately 3.3% of the world's population) are chronically infected with the hepatitis C virus, which causes more than 350 thousands deaths every year [1].
The aim of the work enzyme immunoassay research was to analyze the markers of viral hepatitis C, made in 2016 in diagnostic laboratory of National Scientific Surgery Center under the name of A.N.Syzganov.
Material and methods
This paper analyzes the research immuno markers of viral hepatitis C, isolated from the blood serum of patients treated and examined in National Scientific Surgery Center under the name of A.N.Syzganov in 2016.
A total of the following number of parameters: anti-HCV total antibodies - 1982, including anti-HCV total antibodies in men - 993 and anti-HCV total antibodies in women - 989. Analyses were performed on the unit «Cobas e 411» "Roche" company ( Japan), which is an automatic analyzer which calculates the level of discrimination on the basis of measurements and Cal1 Cal2, outstanding results in the form of discriminatory level index (DLI), cutoff index - COI - sample the signal to discriminatory terms. Thus DLI<1,0 samples evaluated as negative and samples with DLI>1,0 considered positive.
Results and discussion
As a result of the review of the results of studying patients sera anti-HCV total antibodies in males of 993 tests positive results were obtained in 75 cases and negative cases amounted to 918; in the study on anti-HCV total antibodies in women from 989 of 63 analyzes the sample were positive and negative - 926. Age of patients studied ranged from 18 to 75 years.
The diagnosis of HCV infection is based on the identification of specific serological indicators. What is important is not only for the differential diagnosis of hepatitis C, but also for determining the stage of the disease performing the prognosis of its course, evaluating the activity of the process and the effect of the antiviral therapy. To a greater extent, the detection of anti-HCV antibodies is used. It is necessary to carry out genodiagnostics, which is based on the definition of HCV RNA. There is a possibility of detecting circulating antigens of the hepatitis C virus [2,3].
Detection of anti-HCV antibodies is performed using a solid-phase enzyme-linked immunosorbent
assay (ELISA), based on a complex of structural and non-structural viral peptides. At the present stage, diagnostic systems are used that are created from recombinant and/or synthetic fragments of structural and nonstructural HCV proteins (C, NS3, NS4 and NS5). The use of the above diagnosticums makes it possible to shorten the time of the initial establishment of anti-HCV in acute infection, significantly increase the sensitivity and specificity of the reaction.
The frequency of detection of anti-HCV among HCV RNA positive blood samples is close to 100%. In the case of chronic infection, antibodies are detected continuously, and following the elimination of the virus, they are retained (primarily anti-C) for 48 years or more. In this regard, the formal presence of anti-HCV does not always indicate the presence of the virus in the body and does not allow to form an opinion on the activity of the process. For the assumption of viral load, HCV replication activity, chronization risk, separation of acute and chronic hepatitis, the duration of the infection process and the degree of liver damage, it is necessary to apply the establishment of a spectrum of antibodies to various HCV peptides, in addition, to monitor their qualitative and quantitative dynamics. That allows us to have indicative information that needs to be confirmed by other methods [4].
The definition of antibodies to each of the HCV antigens is characterized by an independent diagnostic value. For example, it is known that anti-core, anti-E and anti-NS3 are detected at the earliest stages of seroconversion, anti-NS4 and anti-NS5, as a rule, appear later. As the infectious process develops, the titers of detectable anti-HCV increase.
According to the literature, it was found that the presence of anti-HCV core in the serum is considered a reliable enough sign of viral replication, since there is evidence that their presence correlates with the presence of specific mRNA in the serum.
An anti HCV core test can be used as a confirmatory test when examining samples with "undetermined" results. Samples of blood sera in which the anti-HCV core is determined in a dilution of 1: 1000 are highly likely to have a virus and are really positive. Samples that are positive at 1: 200 dilution and negative at 1: 1000 dilution should be further examined for the presence of viral RNA. With a high probability, these samples can be considered really negative. So, applying a preliminary anti-HCV core test, it is possible to significantly reduce the number of samples that need to be confirmed by polymerase chain reaction (PCR).
There is evidence that in pregnant women (4 weeks of pregnancy and above) a false-positive test for anti-HCV core is possible.
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ВЕСТНИК ХИРУРГИИ КАЗАХСТАНА № 3-2017
Anti-NS3 may be an independent diagnostic indicator of acute viral hepatitis C. High titers of anti-NS3 in acute HCV infection indicate a large viral load, and prolonged persistence of them in the acute phase causes a significant probability of developing a chronic infection process [5].
The detection of anti-NS4 and high titers of these antibodies often talk about the extent of the infectious process, and, apparently, are comparable to the degree of liver damage.
There is evidence that the definition of anti-NS5 in high titers is often indicative of the presence of viral RNA, and in the acute stage appears to be a predictor of the development of a chronic infectious process. In addition, there is an increase in the fact that in the NS5 region of the hepatitis C virus there are sequences that interact nonspecifically with antibodies in human serum samples. With the help of synthetic peptides, epitope mapping of this region was carried out, and sequences were found that specifically interacted with antibodies synthesized specifically to the hepatitis C virus.
The definition of anti-E1/E2 is an important marker for the clinic. Detection of anti-E within 1 month from the onset of the disease and a rapid subsequent decrease may indicate the purification of the body of viral RNA, the determination of anti-E in chronic HCV infection indicates active viral replication.
For many viral infections, such as hepatitis B, hepatitis D, hepatitis A, the detection of IgM antibodies to different viral parameters is an important diagnostic marker of acute or chronic infection. The significance of the detection of antibodies of this class in viral hepatitis C remains not sufficiently elucidated. Different scientists have identified contradictory data. In contrast to the expectations of IgM, the response in the acute phase of HCV infection does not follow the classical pathway of antibody formation: IgM anti-HCV can be detected simultaneously and even later than anti-HCV IgG class. In this regard, the detection of IgM anti-HCV can not be used as an indicator of acute HCV infection. At the same time, the length of the circulation of anticorrosion IgM (35 months) serves as a parameter predicting a persistent infection, and their appearance in chronic hepatitis C indicates the reactivation of the virus, i.e. About the aggravation of the process. This fact is correlated with the rate of decrease in the anticancer IgM and the effectiveness of antiviral therapy.
Immunobloting is based on the differentiated detection of antibodies to various antigens (C, NS3, NS4 and NS5), which, in the form of discrete fractions ("spots"), are fixed on nitrocellulose strips and, after incubation with the serum studied, are labeled Antibodies as well as in the
"usual" ELISA. The result is considered positive when antibodies to two or more antigens are detected [6].
However, without looking at high specificity, modern ELISA systems are not guaranteed from overdiagnosis, i.e. From false positives. To avoid this, a dynamic assessment based on the detection of anti-HCV against discrete antigens (confirmatory, or confirmatory tests) is also required.
Simultaneously with false positive, false-negative results are possible, when anti-HCV antibodies can not be determined despite the presence of the virus in the body. At least three similar situations are known:
1) the initial period of the disease (seronegative phase of acute hepatitis phase "window"),
2) patients receiving immunosuppressants (they can be long-term infected without seroconver-sion),
3) infection with some HCV genotypes (especially 3 and 4).
To date, commercial tests for the detection of antibodies to the hepatitis C virus are used as antigens recombinant proteins or synthetic pep-tides derived from the amino acid sequence of the hepatitis C virus of the 1 st genotype. But there is a growing number of observations that such tests may not be effective enough to reliably detect antibodies when infected with a virus of another genotype. Thus, for example, antigens isolated from the NS3 region of the HCV genotype 1 were found to be 5 times more effective in detecting antibodies in the sera of patients infected with the HCV virus of the 1st genotype than in sera from patients infected with the 2nd or 3rd genotype virus. Studies carried out by a certain number of authors [5] have established the inability of several commercial tests to adequately detect antibodies to hepatitis C virus in patients infected with virus 1a and 3a types. This information is of great importance both for regions where viruses of other types prevail, and for individual populations.
That is, for effective diagnosis of hepatitis C, a highly sensitive test is needed that can adequately react with anti-HCV of any genotype.
In addition to direct investigation of the structure of RNA, data on infection with a specific genotype can be obtained from the study of antibodies against genotype-specific epitopes localized in NS5, NS4 and C proteins of the virus. The list of serotyped variants of HCV at the present moment includes genotypes (subtypes) 1a, 1b, 2a, 2b, 3a and 4a. Data of serotyping and RNA typing coincide in 50-95% of cases, depending on the genotype. Commercial production of diagnostic kits for serotyping in the plate version of enzyme immunoassay and immunoblot is underway.
References
1. Ershov F.I., Romantsov M.G. Viral hepatitis // Medicines used in viral diseases. - MA - 2007 - p. 84-106.
2. Sologub T.V., Romantsov M.G., Ershov F.I. The effectiveness of immunomodulators in the treatment of chronic viral hepatitis //// Medicines used in viral diseases. - M., - 2007. - p. 158-163
3. Sukhanov D.S. Antioxidant Activity remaxol model drug liver damage // Bulletin of St. Petersburg State Medical Academy. Mechnikov. 2008. № 4. p. 127-132.
4. Yushchuk N.D., Klimova E.A., Znoyko O.O., Karetkina G.N., Maksimov S.L., Maiev I.V. Viral
hepatitis: clinical features, diagnosis, treatment. - M., "Geotar-Media" - 2014.- 370 p.
5. Sulkowski MS, Poordad F, Manns MS, Brono-wicki JP, Reddy KR, Harrison SA, et al. Anemia during treatment with peginterferon alfa-2b/ ribavirin with or without boceprevir is associated with higher SVR rates: analysis of previously untreated and previous-treatment-failure patients. J Hepatol 2011;54(suppl 1):S195-S196.
6. Jacobson IM, Catlett I, Marcellin P, Bzowej NH, Muir AT, Adda N, et al. Telaprevir substantially improves SVR rates across all IL28b genotypes in the advanced trial. J Hepatol 2011;54:S1369.
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