Assignment of the group A rotavirus NSP4 gene into genotypes using a hemi-nested multiplex PCR assay: a rapid and reproducible assay for strain surveillance studies.
The rotavirus non-structural protein NSP4 has been implicated in a number of biological functions during the rotavirus cellular cycle and pathogenesis, and has been addressed as a target for vaccine development. The NSP4 gene has been classified into six genotypes (A-F). A semi-nested triplex PCR was developed for genotyping the major human NSP4 genotypes (A-C), which are common in human rotavirus strains but are also shared among most mammalian rotavirus strains. A total of 192 previously characterized human strains representing numerous G and P type specificities (such as G1P[8], G1P[4], G2P[4], G3P[3], G3P[8], G3P[9], G4P[6], G4P[8], G6P[4], G6P[9], G6P[14], G8P[10], G8P[14], G9P[8], G9P[11], G10P[11], G12P[6] and G12P[8]) were tested for NSP4 specificity by the collaborating laboratories. An additional 35 animal strains, including the reference laboratory strains SA11 (simian, G3P[2]), NCDV (bovine, G6P[1]), K9 and CU-1 (canine, G3P[3]), together with 31 field isolates (canine, G3P[3]; feline, G3P[9]; porcine, G2P[23], G3P[6], G4P[6], G5P[6], G5P[7], G5P[26], G5P[27], G9P[6] and G9P[7]) were also successfully NSP4-typed. Four human G3P[9] strains and one feline G3P[9] strain were found to possess an NSP4 A genotype, instead of NSP4 C, suggesting a reassortment event between heterologous strains. Routine NSP4 genotyping may help to determine the genomic constellation of rotaviruses of man and livestock, and identify interspecies transmission of heterologous strains.
Bányai
Krisztián
K
Veterinary Medical Research Institute, Hungarian Academy of Sciences, Budapest, Hungary.|Regional Laboratory of Virology, Baranya County Institute of State Public Health Service, Pécs, Hungary.
Bogdán
Ágnes
Á
Regional Laboratory of Virology, Baranya County Institute of State Public Health Service, Pécs, Hungary.
Szücs
György
G
Regional Laboratory of Virology, Baranya County Institute of State Public Health Service, Pécs, Hungary.
Arista
Serenella
S
Department of Hygiene and Microbiology, University of Palermo, Palermo, Italy.
De Grazia
Simona
S
Department of Hygiene and Microbiology, University of Palermo, Palermo, Italy.
Kang
Gagandeep
G
Department of Gastrointestinal Sciences, Christian Medical College and Hospital, Vellore, India.
Banerjee
Indrani
I
Department of Gastrointestinal Sciences, Christian Medical College and Hospital, Vellore, India.
Iturriza-Gómara
Miren
M
Enteric Virus Unit, Enteric, Respiratory and Neurological Virus Laboratory, Health Protection Agency, London, UK.
Buonavoglia
Canio
C
Department of Public Health and Zootecnic, Faculty of Veterinary Medicine, University of Bari, Bari, Italy.
Martella
Vito
V
Department of Public Health and Zootecnic, Faculty of Veterinary Medicine, University of Bari, Bari, Italy.
D000818:Animals; D001483:Base Sequence; D002415:Cats; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry; D004285:Dogs; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000821:virology; D005838:Genotype; D006023:Glycoproteins / Q000235:genetics*; D000882:Haplorhini; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012401:Rotavirus / Q000145:classification* / Q000235:genetics; D013045:Species Specificity; D013552:Swine; D014118:Toxins, Biological / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D012367:RNA, Viral; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins