Simultaneous Detection and Genotyping of “Norwalk-Like Viruses” by Oligonucleotide Array in a Reverse Line Blot Hybridization Format

摘要: “Norwalk-like viruses” (NLVs) are the most common cause of outbreaks nonbacterial gastroenteritis worldwide. To date, method widely used for typing NLV strains is sequencing and subsequent phylogenetic analysis reverse transcription (RT)-PCR products, which has revealed existence stable distinct lineages (genotypes). This rather costly, not routinely in clinical laboratories, very suitable large numbers samples. Therefore, we have developed a rapid simple genotyping NLVs. The method, designated line blot hybridization, based on nucleotide divergence region gene RNA polymerase can be to classify NLVs into genotypes. was amplified by RT-PCR then hybridized 18 different membrane-bound oligonucleotides that were able discriminate among 13 Application panel 132 positive stool samples from 34 20 sporadic cases collected 6-year period (1994 1999) resulted successful 124 (94%), as confirmed analysis. sequences remaning eight (6%) three did cluster with known Phylogenetic complete partial open reading frame 2 (capsid gene) these one novel genotype (Alphatron) potentially (Amsterdam). allows simultaneous detection NLVs, useful diagnosis obtained large-scale epidemiological studies.