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 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.
Rodríguez-Díaz
Jesús
J
Division of Molecular Virology, School of Medicine, Linköping University, Linköping, Sweden. jerodrig@ivic.ve, chuso333@hotmail.com
Rubilar-Abreu
Elba
E
Spitzner
Markus
M
Hedlund
Kjell-Olof
KO
Liprandi
Ferdinando
F
Svensson
Lennart
L
D000293:Adolescent; D000328:Adult; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D002648:Child; D017931:DNA Primers; D005838:Genotype; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012401:Rotavirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D012400:Rotavirus Infections / Q000821:virology*; D016415:Sequence Alignment; D013548:Sweden; D014118:Toxins, Biological / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D014118:Toxins, Biological; C061332:VP6 protein, Rotavirus; D017361:Viral Nonstructural Proteins