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.
Subject(s)
Phylogeny , Rotavirus Infections/genetics , Rotavirus/genetics , Capsid Proteins/genetics , Capsid Proteins/isolation & purification , Diarrhea/genetics , Diarrhea/virology , Feces/virology , Genotype , Humans , Multiplex Polymerase Chain Reaction/methods , Rotavirus/isolation & purification , Rotavirus Infections/virology , SerotypingABSTRACT
Drasl gene was mapped by in situ hybridization to polytene chromosomes of several sibling species of the Drosophila virilis group and hybrids between them. A 1037 bp fragment of the Drasl gene of the D. virilis genome was used as a probe. The gene sequence is localized to the region of the disk 25 A-B on the chromosome 2 of the polytene chromosome map of D. virilis.
