Analysis of the domestic market for COVID-19 diagnostic kits by real-time reverse-transcription polymerase chain reaction.

COVID-19 is a disease caused by the new coronavirus SARS-CoV-2. Outbreaks were first reported in China on December 31, 2019. Exactly one month later, the WHO declared the outbreak a public health emergency of international concern, and on March 11, it was declared a pandemic.

Development of a reagent kit for the qualitative detection of SARS-CoV-2 virus RNA in naso- and oropharyngeal swabs by real-time RT-PCR.

The coronavirus infection continues to spread around the world. In this regard, the purpose of this work was: to develop a set of reagents for the qualitative detection of SARS-CoV-2 virus RNA. The set was developed by CJSC «Ecolab», 20 positive samples were used to develop the kit.

Development of a PCR assay for the detection of human herpes virus type 7.

A PCR assay has been developed to identify the DNA of the human herpes virus type 7. The search and selection of conserved regions was carried out by comparing the whole genome nucleotide sequences of HHV-7. A fragment duplicated in the HHV-7 genomes was chosen as a target for amplification.

Detection of Y-chromosome marker in plasma of pregnant women using real time PCR: diagnostic accuracy depending on gestation age.

The evaluation of the clinical significance of the test for the detection of the Y-chromosome marker in the plasma of a pregnant woman at different stages of pregnancy by real-time PCR was carried out. The blood samples of 4616 women at 4 to 32 gestation weeks were studied. Identification of the Y-chromosome marker was carried out based on the amplification of a region of the TSPY gene.

Features of the humoral response to immunization "Gam-COVID-Vac" and in patients with COVID-19.

The paper present the results of a survey of people who have undergone immunization with a combined vector vaccine for the prevention of coronavirus infection COVID-19 «Sputnik V - Gam-COVID-Vac», as well as COVID-19 recovalents. Using a quantitative enzyme-linked immunosorbent assay, the levels of specific IgG were determined in persons who had had different degrees of severity before vaccination, in persons who were immuno-negative before immunization, as well as in convalescents who had undergone coronavirus infection of varying severity. The immunological targeting of antibodies against various SARS-CoV-2 proteins is considered.

About quantitative determination of the D-dimer in the blood by immunochromatographic method.

The paper presents the results of the development of a technology for the quantitative determination of D-dimer in blood with an immunochromatographic (LFIA) kit of reagents «LFIA-D-dimer» and instrument accounting of the results. Registration and processing of the digitized indicator of the intensity of staining of the LFIA-test strip using the LFIA-analyzer software allows quantifying the D-dimer content in the sample (in ng DDU/ml). The effectiveness of the proposed approach was evaluated on 258 clinical samples examined in the LFIA with visual and instrument accounting of the results, in comparison with the indicators of D-dimer determination in ELISA.

Assessment of the state of intestinal microbiocenosis based on bacterial endotoxin and plasmalogen in elderly persons with type 2 diabetes mellitus pathology.

The concentration of bacterial plasmalogen 18a and endotoxin in the blood of elderly people 45-90 years old with the pathology of type 2 diabetes mellitus (DM 2) - the main group and without diabetes mellitus - the comparison group was investigated. The concentration of both plasmalogen 18a and endotoxin in the blood of individuals with DM 2 pathology is statistically significantly higher than in the blood of individuals without DM 2 pathology. To assess the state of microbiocenosis and predict type 2 diabetes mellitus, decisive rules have been determined in the form of threshold values of plasma concentrations 18a and endotoxin in the blood of elderly people with a suspected or established diagnosis of type 2 diabetes.

Development of test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot» format.

Test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot» format has been developed. A preliminary study of diagnostic effectivity on clinical samples of blood serum from patients with COVID-19 and healthy donors showed its high sensitivity and specificity. The new test kit allows to detect IgM to all four structural antigens of SARS-CoV-2 and can be used as a confirmatory test to verify indeterminant screening results in laboratory etiological diagnosis of COVID-19.

Development of immunofluorescent diagnostics for the determination of IgM and IgG antibodies to herpes simplex virus types 1 and 2.

The Russian kits «HSV-1-Fluorogen-screen» and «HSV-2-Fluorogen-screen» have been developed for the determination of antibodies M and G to herpes simplex virus types 1 and 2 by the immunofluorescence reaction. The kits were used to examine the positive and negative standard «EKOlab» panels sera and showed 100% sensitivity and specificity of the developed tests. 125 samples of blood serum from people with clinical diagnoses such as herpetic, cytomegalovirus infections, pyelonephritis, conjunctivitis and central nervous system damage were tested in parallel with using the enzyme-linked immunosorbent assay systems from different manufacturers and the developed tests «HSV-1-Fluorogen-screen» and «HSV-2-Fluorogen-screen».

Immune blotting as a method for diagnosis of toxoplasmosis.

Test kit for detection of IgG-antibodies to individual antigens of Toxoplasma gondii by immune blotting («Western blot» format) has been developed. Laboratory testing with first international WHO standard «Anti-toxoplasma serum (IgG), human, Lyophilized, 20 IU / ampoule» (NIBSC, Great Britain) demonstrated the analytical sensitivity of the new kit equal to 10 IU / ml. Study of the diagnostic efficiency of the new kit showed its high sensitivity, equal to 98.

Immunochromatographic test for differentiation detection of IgM and IgG to SARS-CoV-2.

The study presents the results of the creation and evaluation of the diagnostic characteristics of the rapid immunochromatographic test for the qualitative detection and differentiation of IgM/IgG antibodies to SARS-CoV-2 in human serum, plasma, and whole blood "ИХА-COVID-19-IgM / IgG". Have been tested some samples without antibodies to SARS-CoV-2 and a samples with two and one type of specific antibodies. The coincidence of the results of immunochromatographic analysis with the results of the immunochemiluminescent method was 87.

Development of an enzyme-linked immunosorbent assay for the detection of IgG-antibodies to the causative agent of COVID-19 in human serum (plasma).

A new original Russian test kit for the detection of IgG-antibodies to the causative agent of COVID-19 - coronavirus SARS-CoV-2 by the method of enzyme-linked immunosorbent assay (ELISA) on a solid-phase «ELISA-SARS-CoV-2-AT-G» has been developed. In comparative tests with similar test systems «Vitrotest® SARS-CoV-2 IgG» (Vitrotest, Ukraine) and «Anti-SARS-Cov-2 ELISA (IgG)» (EUROIMMUN AG, Germany) high diagnostic efficiency of the new test system was shown.

[Representation of the results when testing for syphilis.].

In medical organizations special forms (approved by orders of the Ministry of health and the Central statistical office) are used to present the results of laboratory examination of the population for syphilis. Currently, these forms include laboratory technologies that do not meet modern standards. In 2018-2019 the normative documents, regulating the use of accounting forms of medical documentation in the public health system, and the practice of using forms with the results of laboratory examination for syphilis were studied in 35 branches of 3 medical organizations of the Lyubertsy district of the Moscow region and in 18 branches of Moscow city was conducted.

[Comparative evaluation of the immunochromatographic test for detection of hemoglobin.].

The evaluating results of the diagnostic characteristics of an immunochromatographic test for the detection of fecal occult blood (hemoglobin) were presents in the article. The test was approved samples without hemoglobin and model samples containing of standardized preparation of hemoglobin in various concentrations in comparison with two immunochromatographic test systems. The developed test system identified hemoglobin in concentrations higher 50 ng/ml in 99,2% cases, the specificity of the analysis was 98,1%.

[Development of a set of reagents for detection of class G immunoglobulins to human herpes virus type 7 by the method of immune blotting in the format «Western blot».].

A new original domestic set of reagents has been developed for the determination of class G immunoglobulins to individual human herpes virus antigens of type 7 by the method of immune blotting in the "Western-blot" format. Preliminary clinical trials were conducted using 134 serums of healthy children aged 1-16 years who underwent diagnostic testing. Stud y of diagnostic efficiency ofthe new kit showed high sensitivity, comparable to the sensitivity of the reaction indirect immunofluorescence and high specificity, which is manifested in the absence of false positive results when testing samples containing immunoglobulin G to herpes virus 6 type.

[Infection with Epstein-Barr virus in certain groups of the population of the Moscow region.].

The serum of 1006 children aged 0-18 years and elderly people aged 60 to 90 years and older for the presence of specific class G immunoglobulins to the Epstein-Barr virus was studied using enzyme immunoassay. The dependence of seropositivity of children on their age and seropositivity of more than 98% of all surveyed elderly people is shown.

[Immunochromatographic test for determination of D-dimer.].

The developing and the testing results of an immunochromatographic test for the D-dimer qualitative determination were presents in the article. The test was approved blood plasma samples in comparison with a quantitative enzyme-linked immunosorbent assay (ELISA). The results of assay with developed test were the same with ELISA results for 87,1% and 100% for samples with increased (more than 400 ng/ml FEU) and normal concentration of D-dimer, respectively.

[Immunochromatographic test for detection of fecal occult blood.].

The developing and evaluating results of the diagnostic characteristics of an immunochromatographic test for the detection of fecal occult blood (hemoglobin) were presents in the article. The test was approved samples without hemoglobin and model samples containing of standardized preparation of hemoglobin in various concentrations. The developed test system identified hemoglobin in concentrations higher 5 μg / g feces in 97.

[First results of application of an enzyme immunoassay in the serological diagnostics of infection caused by herpesvirus type 7.].

HHV-7 (Human Herpes virus type 7) is a relatively recently discovered, ubiquitous beta herpes virus. In Russia, the diagnosis of HHV-7 infection is carried out by PCR, which determines the DNA in serum or plasma, if there are clinical indications. Laboratory serological tests for HHV-7 are not performed.

[Current state and development trends of domestic medical device market for in vitro diagnostics in the segment of diagnostic reagents and their kits.].

The problem of import substitution in the Russian medical device market for in vitro diagnostics (MD) has not been solved, despite a number of measures taken by the government of the Russian Federation since 2011. The reasons hampering this process are the lack of modern analytical equipment of domestic production and deficiencies of the current regulatory system for MD circulation in the market. It is proposed the transition from licensing system of MD registration to notification system as a radical measure to solve the problem of import substitution.

[Standardization of component manufacturing conditions and setting up the immunofluorescence reaction on the example of syphilis laboratory diagnostics (literature review).].

The use of technology for preparing components for immunofluorescence reaction (RIF) in a number of clinical diagnostic laboratories (CDL) in accordance with the recommendations of the Guidelines introduced by order of the RF Ministry of Health No 87 dated March 26, 2001 does not guarantee the required standardization of RIF preparation and conduct conditions. The latter is achieved by using the appropriate industrial production reagent kits in the СDL practice.

[Application of indirect enzyme immunoassay to detect specific IgM for human herpes virus type 6.].

In recent years, timely diagnosis of human herpes virus infection of type 6 (HHV6), especially in pediatrics, has acquired particular relevance. In the spectrum of serological diagnostics, an indirect ELISA method was used to detect IgM against HHV-6 in a study of samples of children sera (198), conditionally healthy patients (148) and donors (145). The use of ELISA IgM HHV6 is more significant for the expansion of the complex of the diagnostic study of the disease associated with primary HHV6 infection or its reactivation.

[Development of a test kit for the detection of IgG-antibodies to individual antigens of Rubella virus by immunoblotting (Western blot).].

A Russian test kit for detecting IgG-antibodies to individual antigens of Rubella virus by immunoblotting (Western Blot) was designed. The new test kit is intended for confirmation of positive screening results and for differentiation of stages of infection. It has sensitivity and specificity not conceding one of its analogues, the «Anti-Rubella virus (IgG) WESTERNBLOT» («EUROIMMUN AG», Germany).

[The synchronous detection of serological markers of main herpes viral human infections].

The new original national set of reagents "Line-Blot human herpesvirus-profle" was developed to detect antibodies (IgG - set 1 and IgM - set 2) to main agents of herpesvirus infections (HSV-1, HSV-2, VZV, EBV, CMV) in the format of linear immunoblotting. The preliminary clinical trials were implemented using 319 samples of blood serum: HIV-infected patients (n=128), pregnant women (n=86) and persons undergoing treatment or diagnostic examination (n=105). In the case of every infection agent analyzes were implemented using enzyme-linked immunosorbent assay and linear immunoblotting with new set and its analogues of German manufacturer.

Multiplex immunoassay for detection of immunoglobulin G to herpes simplex virus types 1, 2 and cytomegalovirus based on PHOSPHAN technology.

We have developed a multiplex immunoassay test (immunochip) based on PHOSPHAN technology for the detection of immunoglobulin G to herpes simplex virus (HSV) types 1, 2 and cytomegalovirus (CMV). The immunochip consists of HSV type specific gG1 (HSV-1) and gG2 (HSV-2) recombinant antigens, the lysate antigen for detection of total IgG to both HSV types (HSV 1/2), and CMV specific chimeric recombinant antigen containing the immunodominant sequences of pp150, gB, pp28 and pp52 proteins. The sensitivity and specificity of simultaneous IgGs detection with recombinant proteins were comparable to the commercial ELISA kits regardless of the kind of investigated serum specimens (patient sera, standard serum panels).