Design of a multiplex nested PCR for genotyping of the NSP4 from group A rotavirus

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 strain...

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Bibliographic Details
Published in:	Journal of virological methods 2008-05, Vol.149 (2), p.240-245
Main Authors:	Rodríguez-Díaz, Jesús, Rubilar-Abreu, Elba, Spitzner, Markus, Hedlund, Kjell-Olof, Liprandi, Ferdinando, Svensson, Lennart
Format:	Article
Language:	eng
Subjects:	Adolescent Adult Antigens, Viral - genetics Base Sequence Biological and medical sciences Capsid Proteins - genetics Child DNA Primers Fundamental and applied biological sciences. Psychology Genotype Genotyping Glycoproteins - genetics Group a rotavirus Humans MEDICIN MEDICINE Microbiology Molecular Sequence Data Multiplex PCR Nested PCR NSP4 Polymerase Chain Reaction - methods Rotavirus Rotavirus - classification Rotavirus - genetics Rotavirus - isolation & purification Rotavirus Infections - virology Sequence Alignment Sweden Techniques used in virology Toxins, Biological - genetics Viral Nonstructural Proteins - genetics Virology
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Summary:	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.
ISSN:	0166-0934 1879-0984 1879-0984