Abstract

A STUDY ON METHOD DEVELOPMENT FOR DETECTION OF DIFFERENT VIRUAL ANTIBODIES AND VIRUS BY USING RT-PCR METHOD

Gowtham Mandadapu, Prachetha Kolli and Kurra Venkata Gopaiah*

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Abstract

Broadly targeted real-time PCR techniques for early diagnosis of viral infection by detection of virus nucleic acid (DNA and RNA) and virus antibodies (IgM) were developed. I. The developed QRT-PCR detected EV RNA from samples which contain a few copies of RNA (1-10 gene equivalents per PCR reaction) and from a broad variety of EVs in reference panels. II. Our results indicate that QPIA achieves a rapid diagnosis of enterovirus infection. The combination of QRT-PCR and QPIA detected 40/43 (93%) of aseptic meningitis cases from which both serum and CSF were available. III. A broadly targeted, simple, single tube real-time degenerated quantitative PCR (QPCR) technique for detection of JCV, BKV and SV40 DNA was developed. At least down to 10-100 gene equivalents of target DNA per PCR reaction were detected. The QPCR is simple to perform and should be valuable for diagnosis of polyoma virus infection. IV. The previous results regarding the frequencies of IgM antibodies against EVs in newly diagnosed T1D children, siblings and controls were confirmed by using viral antigen isolated from T1D cases at onset (T1D-EV-QPIA).

Keywords: PCR, DNA, RNA, QRT-PCR, QPIA, CSF, JCV, BKV, SV40, T1D, EVs.