[Evaluation of real-time multiplex PCR effectiveness for group a rotavirus genotyping].

ABSTRACT

AIM: Evaluate resolution and diagnostic significance of real-time multiplex PCR (MP RT-PCR) as a platform for group A rotavirus G/P genotyping test-systems. MATERIALS AND METHODS: Primer and DNA probe construction for an experimental test-system based on MP RT-PCR was carried out by using specialized PC programs and sequence databases GenBank NCBI, EMBL Nucleotide Sequence Database etc. The experimental genotyping test-system was tested using 116 clinical samples with confirmed rotavirus infection and 14 biosamples negative for group A rotavirus RNA. Selective sequencing of VP7, VP6, VP4 gene mark-erloci was carried out as a reference method forverifying determination of rotavirus genotype. RESULTS: Specific interaction between primers and DNA probes with genotype-specific loci of retrovirus genome segments and a lack of false-negative signals, complete match ofgenotyping results obtained by MR RT-PCR and sequenc- ing methods were established. CONCLUSION: The resolution of MP RT-PCR methods allows designing test-systems that can confidently identify rotavirus genotypes with effectiveness of 90% and above.