Designing and validation of new primers for efficient genotyping of animal G3 rotaviruses

作者：Malik Yashpal S^*; Sharma Kuldeep; Vaid Nirupama; Batra Munish; Prasad Minakshi; Prasad Gaya; Gulati Baldev R; Bhilegaonkar Kiran N

来源：Veterinarski Arhiv, 2013, 83(6): 611-622.

• group A rotavirus
• animals
• G3
• G6
• G10 genotype
• VP7 gene
• genetic diversity
• multiplex-RT-PCR
• genotyping
• G3 primers

The present study describes the problem of genotyping failures of animal rotaviruses with existing and in-use G3 genotyping primers. To overcome the problem, published and in-use 03 typing primers with sequences of VP7 genotyping regions from human and animal G3 rotavirus isolates were evaluated. The sequence alignment analysis showed that existing in-use 03 primers exhibit higher complementarities with rotavirus isolates of G6, G8 and G10 genotype specificities. The existing G3 primers showed up to 9 nucleotide mismatches with the animal origin rotavirus isolates of G3 genotype specificity. The modified G3 genotyping primers yielded positive amplification in all the G3 isolates of animal origin, with no incorrect amplification with any other group A rotavirus genotypes viz. G6 and G10. We advise the use of the proposed primers in molecular surveillance studies to discover the truly dominant genotypes of rotaviruses in animals.

• 出版日期2013-12