[The identification of viruses of human papilloma of high carcinogenic risk and evaluation of physical status of viral DNA using technique of polymerase-chain reaction under affection of cervical epithelium].

The DNA of virus of human papilloma of high carcinogenic risk was detected in 116 cervical samples. At that, the morphological symptoms of background processes are detected in 19 samples, CIN 1 in 9, CIN 2 in 23, CIN 3 in 54 (and out of them carcinoma in situ in 13), epidermoid cancer (squamous cell carcinoma) in 11 cases. The viral load of human papilloma of high carcinogenic risk in all samples of DNA exceeded threshold of clinical value (3 lg copies of DNA of human papilloma/105 cells). The genetic typing of human papilloma of high carcinogenic risk revealed the dominance of human papilloma of type 16 in 49.7%, type 33 in 15.3%, type 31 in 12.3% and type 45 in 5.5%. In women with background processes in cervix of the uterus DNA of human papilloma type 16 was detected more often in episome form. In case of dysplastic alterations of epithelium and cervical cancer DNA of human papilloma type 16 is detected in mixt form with different degree of integration into cell genome.

[The first experience of application of standardized genotype technique of identification of HIV-tropism].

The antagonists of co-receptor CCR5 are an ultimately new class of preparations to treat HIV-infection. The mechanism of action of the preparations of this class is in the selective binding with co-receptor CCR5. This process results in the prevention of penetration of HIV into cell. Before prescribing the CCR5 antagonists the detection of viral tropism has to be done. Recently, in Russia the genotype technique of tropism detection was registered which can be used in clinical practice. The present article describes first experience of application of the given technique to clinical samples. The high correlation was established while comparing with the results of reference laboratory.

[Detection of rubella virus RNA in clinical material by real time polymerase chain reaction method].

AIM: Development of a reagent kit for detection of rubella virus RNA in clinical material by PCR-RT. MATERIALS AND METHODS: During development and determination of analytical specificity and sensitivity DNA and RNA of 33 different microorganisms including 4 rubella strains were used. Comparison of analytical sensitivity of virological and molecular-biological methods was performed by using rubella virus strains Wistar RA 27/3, M-33, "Orlov", Judith. Evaluation of diagnostic informativity of rubella virus RNAisolation in various clinical material by PCR-RT method was performed in comparison with determination of virus specific serum antibodies by enzyme immunoassay. RESULTS: A reagent kit for the detection of rubella virus RNA in clinical material by PCR-RT was developed. Analytical specificity was 100%, analytical sensitivity - 400 virus RNA copies per ml. Analytical sensitivity of the developed technique exceeds analytical sensitivity of the Vero E6 cell culture infection method in studies of rubella virus strains Wistar RA 27/3 and "Orlov" by 11g and 31g, and for M-33 and Judith strains is analogous. Diagnostic specificity is 100%. Diagnostic specificity for testing samples obtained within 5 days of rash onset: for peripheral blood sera - 20.9%, saliva - 92.5%, nasopharyngeal swabs - 70.1%, saliva and nasopharyngeal swabs - 97%. Positive and negative predictive values of the results were shown depending on the type of clinical material tested. CONCLUSION: Application of reagent kit will allow to increase rubella diagnostics effectiveness at the early stages of infectious process development, timely and qualitatively perform differential diagnostics of exanthema diseases, support tactics of anti-epidemic regime.

Cellular expression of INK4a gene in cells of bladder cancer associated with human papilloma virus-16.

Hyperexpression of p16(INK4a) protein is an early marker of cervical cancer. Hyperexpression of INK4a gene encoding this protein at the level of mRNA and p16(INK4a) was detected in tumor cells of some patients with bladder cancer associated with human papilloma virus-16. However, in contrast to cervical cancer, this phenomenon in urothelial carcinomas does not correlate with expression of human papilloma virus-16 oncogenes E6 and E7.

[Are human papillomaviruses responsible for the occurrence of bladder cancer].

A female patient with recurrent bladder cancer underwent complex examination. The primary tumor removed in 2004 showed human papillomavirus (HPV) 16 DNA, mRNA corresponding to HPV16 oncogene E7, as well as HPV16 protein E7. The patient is a smoker who has been working at a chemical factory for over 20 years. During tumor recurrence in 2009, there was no DNA of high-risk HPV types in the cancer cells. HPV16 E7protein and cellular p 16(INK4alpha), an indicator of HPV-induced carcinogenesis, were not found. Colposcopy revealed no precancerous changes in the epithelium of the cervix uteri. The cervical epitheliocytes contained no high-risk HPV DNA, E7 and p16(INK4alpha) proteins. It seems expedient to continue in vitro studies of the possible role of HPV in urothelial carcinogenesis on an experimental model.

[Infection with high-risk human papillomavirus and prognosis of cervical carcinoma].

Real-time polymerase chain reaction procedure was used to evaluate bioptic tumor samples from patients suffering cervical carcinoma (CC) stages I-IV. Out of 110 patients, high-risk human papillomavirus (HPV) infection was identified in 98 (89.1%), HPV type 16--63, HPV type 18--10 and HPV type 45--5. One of genotypes 31.33, 35, 39, 52, 58, 59 was established in 8 and a combination of several genotypes of the virus--12 patients. Frequency of remission in CC patients associated with HPV type 16 who had survived 3 years was significantly higher than in the same category associated with HPV type 18 (p=0.03). Relapse frequency and mortality rates in patients with tumors associated with one of viruses 31.33, 35, 39, 52, 58 or 59 were higher as compared with HPV type 16--associated cases 2 years (p=0.03) or 3 years on (p=0.11), respectively. A similar trend was established for squamous-cell tumors stages 1 and 2 (p=0.07) (p=0.12), respectively. No difference was observed in efficacy of therapy for infection with one or a combination of several genotypes of high-risk HPV. Hence, the genotype of virus is believed to be a factor of prognosis in CC early cancers. However, a definitive conclusion cannot be reached until results of a larger body of evidence and longer follow-up are available.

[Detection of oncoprotein E7 HPV16 in the cancer and normal urinary bladder urothelium].

Oncoprotein E7 HPV16 was detected by immunohistochemical staining with specific polyclonal antiserum [Fiedler et al., 2004] in 7 out of the 24 (29.2%) studied bladder cancer specimens. The result is in good agreement with the hypothesis that HPVs take part in the carcinogenesis of the urothelium. However, some of the observations made seem rather hard to be interpreted at present. The latter include the detection of E7 HPV16 in a small number of cancer cells in a few bladder cancer specimens being examined; the presence of this protein in the cytoplasm, rather in the cancer cell nuclei, and its detection in some morphologically normal bladder urothelial specimens from non-cancer patients. Thus, the hypothesis that HPVs are implicated in the carcinogenesis of the bladder urothelium deserves further verification.