Vol 49, No 1 (2004)

We analyzed 52 blood samples obtained independently from among individuals, who had never practiced the intravenous administration of drugs, for the purpose of detecting the subtypes of HIV-1 that circulated during 19992002. The study materials were analyzed by the methods of gag/env heteroduplex mobility assay and by env gp120 C2-V3 sequencing. Four viral subtypes (A,B,C and G) and a recombinant gagA/envB were detected in subjects contaminated through heterosexual contacts. Noteworthily, HIV-1 variations of subtype A, which were found in 22 (73.3%) of 30 analyzed samples, were predominant in this risk group. An analysis of nucleotide sequences exposed a high degree of homology between the viruses, detected previously among drug-addicts, and the isolates detected by us in subjects contaminated heterosexually. However, HIV-1, subtype B, detected by us in all 16 studied cases, still continue to circulate among the males infected through homosexual contacts with men.

HBs antigens were subtyped in blood samples of intravenous drug-addicts (IDA) and of donors from as many as 10 cities of Western Siberia by using the immune-enzyme assay with 6 high-specific monoclonal antibodies. Two HBsAg subtypes were found, in a ratio of 3% ayw2: 97% ayw3 (varA and varB), in IDA blood samples from Novosobirsk. Three HBsAg subtypes were found, in a ratio of 57% ayw2: 42% ayw3 (varA and varB): 1% adw2, in the donors' blood samples. The obtained data are sufficient for developing the first national sera panel containing different HBsAg subtypes of hepatitis В virus typical of Russia.

Polymerase chain reaction (PCR), that can amplify a fragment of the DNA-polymerase gene of 4 herpes viruses, i.e. herpes simplex viruses, type 1 (HSV-1), herpes simplex viruses, type 2 (HSV-2), Epstein-Barr virus and cytomegalovirus , was made use of to study the genetic polymorphism of HSV-1 and HSV-2 strains. The obtained amplicons were analyzed by the method of restriction-size fragments' polymorphism (RSFP) with restrictases Rsal, Taql and Hinfl. Four HSV-1 strains had an identical restriction profile. Strain G (HSV-2) also displayed the expected restriction profiles, however, contradictory results were obtained for strain BH (HSV-2): the restriction profiles with restrictases Hinfl and Rsal corresponded to HSV-2, and the restriction profile with Taql corresponded to HSV-1. The sequencing of appropriate fragments of strains G and BH revealed a dot-type mutation localized in Taql restriction site. The thus worked out PCR was used jointly with RSFP in the genoty ping of 75 urogenital samples obtained from women with genital herpes who were treated at Moscow patient-care facilities. HSV-1 and HSV2 were detected in 18 (24%) and 57 (76%) of samples, respectively. No changes were registered in the restriction profile for HSV-2 among the investigated samples and all of them had the restriction profile similar to that of strain G. The conclusion is that genital herpes associated with HSV-2 is genetically stable within its Moscow population.

The conducted virologic, serological and molecular-and-biological investigations showed that virus Coxsakie B4, isolated from cerebrospinal fluid and rhinopharyngeal lavages of patients, was the main etiological agent, which caused mainly an outbreak of enterovirus infection in the city of Vitebsk in 2001. Coxsakie B4 viruses were found in 30% of samples by using the cultural method and in 76.9% of samples - by polymerase chain reaction (PCR) while carrying out the sanitary-and-virologic investigations in drinking water, including in infection foci. Besides, we found infectious non-cytolitic enteroviruses (EV) with changed biological properties (which could not be detected by the classic cultural method) in drinking water by using the method of PCR integrated with cell culture in the "-" strand of EV RNA. Peculiar clinical-and-epidemiological characteristics of the disease outbreak, i.e. "explosive" onset, multiple clinical forms, mixed EV infections and disease decay after drinking-water chlorination, as well as the isolation of one and the same EV serotype from patients and from water testify to a possible water origin of the outbreak.

Bovine leucosis virus (BLV), circulating in the Ukrainian territory, was characterized through the definition of its subspecies affiliation. The pro-viral BLV DNA was isolated from peripheral-blood lymphocytes of naturally-HIVinfected black-variegate animals taken from leucosis-affected farms in the Kharkov Region. The env-gene fragment of pro-viral DNA was amplified, sequenced and analyzed after the amplicon had been treated by three restriction enzymes, i.e. BamH I, Bel I and Pvu II. According to the analysis of restriction-fragments' length polymorphism, the Ukrainian BLV isolate can be classified as belonging to the Australian subspecies, i.e. to one of the 3 known subspecies. Multiple alignment and phylogenetic analysis of the env-gene fragment of BLV isolates from the EMBL database showed that evolutionally the Ukrainian isolate is distantly located from the isolates' clusters of the Belgian, Japanese and Australian subspecie and has the biggest quantity (4) of non-coinciding nucleotides for the analyzed highly conservative locus of the BLV env-gene with a length of 444 pair of nucleotides.