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. The performance of the assay was tested on a synthetic matrix and clinical samples. The developed assay has high sensitivity and specificity and showed good efficiency in detecting HHV-7 DNA in clinical samples.

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-screen¼ and «HSV-2-Fluorogen-screen¼ 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 «EKOlab¼ 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-screen¼ and «HSV-2-Fluorogen-screen¼. A high degree of matching of results with comparison sets was observed in examined samples. The developed diagnostics can be successfully used in clinical practice both for screening and for verification of results of the diagnosis of herpesvirus infections caused by herpes simplex virus types 1 and 2.

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 87.2%. Test kit can be use as the rapid diagnostic test in the context of the COVID-19 pandemic and to assess the immune status of convalescents.

[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. Currently, in the scientific literature, information on clinical studies of HHV-7 infection, its epidemiology, and its role in the etiopathogenesis of various diseases is insufficient. This article presents the results of the development of a test kit "ELISA-HHV-7-IgG", designed to detect specific IgG to HHV-7 by enzyme-linked immunosorbent assay (ELISA). Studies of the sensitivity and specificity of the developed test kit, including potential cross-reactive samples positive for other herpesvirus infections, were carried out. In the context of serological diagnostics, we studied samples of the sera of children (n = 167) and old people (n = 238) for the presence of IgG to HHV-7 with confirmation of the results in the indirect immunofluorescence reaction and the immune blotting reaction. The research results are evidence of the possibility of using the developed test kit for serological diagnosis of HHV-7 infection.

[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.

[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. The indices of diagnostic informativeness were calculated according GOST R 53022.3-2008. The obtained data permitted to begin registration in the Russian Federation the new set of reagents intended to synchronous screening of monospecifc antibodies to main agents of human herpesviral infections and establishment of activity of manifestation of infection and time of infection by some of them.