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Determination of Porcine Rotavirus Serotypes by RT-PCR and RFLP Analysis
Author(s) -
Hong-Gi Min,
Yong-Hwan Lim,
ShienYoung Kang
Publication year - 2006
Publication title -
journal of bacteriology and virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.179
H-Index - 12
eISSN - 2093-0429
pISSN - 1598-2467
DOI - 10.4167/jbv.2006.36.4.255
Subject(s) - restriction enzyme , restriction fragment length polymorphism , typing , biology , serotype , feces , terminal restriction fragment length polymorphism , rotavirus , microbiology and biotechnology , veterinary medicine , polymerase chain reaction , virology , genetics , gene , virus , medicine
G and P tying of group A porcine rotaviruses (PoRV) from field fecal samples were performed using reversetranscriptase polymerization chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) analysis. After amplifying full length VP7 and partial length VP4 genes, restriction endonucleases were used to digest and analyze the cutting pattern of the gene products. After analysis of digests with restriction endonucleases, seven and six RFLP types were observed for VP7 and VP4, respectively. The G typing analysis of 50 fecal samples revealed that 68% (34/50) were G4, which included G4-like (22/50); 22% (11/50) were G5; 6% (3/50) were G4 and G5 mixed types. The P typing analysis of the same fecal samples revealed that 36% (18/50) were P2B, 52% (26/50) were P9, 1 sample (2%) was a mixture of P2B and P9. Combinations of G and P types, the G4P2B and G4P9 types including G4-like accounted for 26% (13/50) and 32% (16/50), respectively. The G5P2B and G5P9 type also represented 4% (2/50) and 18% (9/50) of the samples. No G3 and G11 or other new P types were identified from the samples tested. Information on the G and P types and G/P combinations in the field fecal samples is useful for developing more effective PoRV vaccines and understanding the epidemiology of PoRV infections in the field.

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