J Bacteriol Virol.  2006 Dec;36(4):255-261. 10.4167/jbv.2006.36.4.255.

Determination of Porcine Rotavirus Serotypes by RT-PCR and RFLP Analysis

Affiliations
  • 1Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea. sykang@chungbuk.ac.kr

Abstract

G and P tying of group A porcine rotaviruses (P(o)RV) 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.

Keyword

Porcine rotaviruses; G type/P type; RT-PCR; RFLP

MeSH Terms

DNA Restriction Enzymes
Epidemiology
Polymerization
Polymers
Polymorphism, Restriction Fragment Length*
Rotavirus*
Vaccines
DNA Restriction Enzymes
Polymers
Vaccines

Figure

  • Figure 1. Restriction endonuclease cleavage patterns of reference PoRV VP7 (A) and VP4 (B) genes. M: Marker, E: EcoRI, B: BamHI, H: HindIII, N: NlaIV, V: VspI, R: EcoRV

  • Figure 2. Restriction endonuclease cleavage patterns of PoRV VP7 genes from fecal samples. M: Marker, E: EcoRI, B: BamHI, H: HindIII, N: NlaIV, V: VspI I, IIa, IIb, IIIa, IIIb, IIIc and IV represent VP7 RFLP type.

  • Figure 3. Restriction endonuclease cleavage patterns of PoRV VP4 genes from fecal samples. M: Marker, E: EcoRI, R: EcoRV, BamHI, H: HindIII 1, 2, 3, 4, 5, and 6 represent VP4 RFLP type.


Reference

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