Veterinary Archives, Vol. 83 No. 6, 2013.
Original scientific paper
Designing and validation of new primers for efficient genotyping of animal G3 rotaviruses.
Yashpal S. Malik
orcid.org/0000-0002-2832-4854
; Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Kuldeep Sharma
; Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Nirupama Vaid
; Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Munish Batra
; GB Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India
Minakshi Prasad
; Animal Biotechnology, CCS Haryana Agricultural University, Haryana, India
Gaya Prasad
; Animal Biotechnology, CCS Haryana Agricultural University, Haryana, India
Baldev R. Gulati
; National Research Center on Equines, Hisar, Haryana, India
Kiran N. Bhilegaonkar
; Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Abstract
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 G3 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 G3 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.
Keywords
group A rotavirus; animals; G3; G6; G10 genotype; VP7 gene; genetic diversity; multiplex-RT-PCR genotyping; G3 primers
Hrčak ID:
111820
URI
Publication date:
9.12.2013.
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