[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.

[The pattern of central nervous system lesions in HIV-infected patients of the specialized unit in infectious disease hospital].

AIM: To define the incidence and features of brain lesion (BL) in HIV-infected inpatients. SUBJECTS AND METHODS: Four hundred and fifty-eight patients with Stage 4B HIV infection (AIDS) and central nervous system (CNS) lesion admitted to Infectious Diseases Hospital Two, Moscow, were followed up in 2003-2009. The authors used cerebrospinal fluid (CSF) microscopic and bacteriological assays for DNA of T. gondii, M. tuberculosis, herpes simplex virus (HSV) types 1 and 2, cytomegalovirus (CMV), HSV type 6, and varicella-zoster virus, Cr. neoformans, C. albicans, C. glabrata, and C. krusei. Blood and CSF were tested for IgM and IgG T. gondii antibodies; brain magnetic resonance imaging was carried out. RESULTS: In patients with late-stage HIV infection, the principal cause of neurological diseases was cerebral toxoplasmosis (34.7% of BL cases) and a generalized process involving the brain, lung, heart, liver, and eyes in 11.5%. There was commonly cerebral toxoplasmosis concurrent with CMV infection with clinical manifestations. 16-32% of the inpatients developed tuberculosis meningoencephalitis that was a manifestation of hematogenous disseminated tuberculosis involving the lung. There was a rise in the incidence of cancers (brain lymphomas, astrocytomas) running with CNS lesion. Mental disorders progressing to dementia were a distinctive property of CMV ventriculoencephalitis, one of the leading factors in the development of AIDS dementia complex. Molecular diagnostic techniques are needed to ascertain the etiology of BL in HIV infection. CONCLUSION: The CSF test for DNA of causative agents is a specific and most sensitive method for diagnosing a relevant CNS lesion.