1. Design of a multiplex nested PCR for genotyping of the NSP4 from group A rotavirus.
- Author
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Rodríguez-Díaz J, Rubilar-Abreu E, Spitzner M, Hedlund KO, Liprandi F, and Svensson L
- Subjects
- Adolescent, Adult, Antigens, Viral genetics, Base Sequence, Capsid Proteins genetics, Child, DNA Primers, Genotype, Humans, Molecular Sequence Data, Rotavirus isolation & purification, Sequence Alignment, Sweden, Glycoproteins genetics, Polymerase Chain Reaction methods, Rotavirus classification, Rotavirus genetics, Rotavirus Infections virology, Toxins, Biological genetics, Viral Nonstructural Proteins genetics
- Abstract
A novel PCR method was developed to discriminate amongst genotypes A-C of the rotavirus non-structural protein 4 (NSP4). Genotype-specific primers were designed that correctly identified the NSP4 genotype when evaluated as a multiplex PCR with cell culture adapted rotavirus strains. Rotavirus strains B223 SGIG6P6[1], NCDV SGIG6P6[1] and SA11 SGIG3P5B[2] were used as control for NSP4 genotype A; A34 SGIG5P14[23], Gottfried SGIIG4P2B[6] and Wa SGIIG1P1A[8] for NSP4 genotype B; RRV SGIG3P5B[3] for NSP4 genotype C. Subsequently, the same set of specific primers was used to genotype a set of 77 Swedish clinical samples. The results showed that all human clinical samples analyzed belong to the NSP4 genotype B and the VP6 subgroup II.
- Published
- 2008
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