A Rapid Method for Extraction of Rotavirus RNA from Fecal Samples and Genotyping of Rotavirus by Reverse Transcription-PCR

Rotavirus is the leading causative agent of severe diarrhea among children.1 The rotavirus genome consists of 11 double-stranded RNA (dsRNA) segments enclosed in a double-shelled capsid. The outer shell is composed of a major glycoprotein (vp7), which defines the rotavirus serotype specificity. Nucleotide sequence analysis of vp7 cDNAs from different isolates indicates the presence of variable regions that are highly conserved within a given serotype, and distinct among different serotypes.2 Gouvea et al.,3 demonstrated a reverse transcription-PCR (RT-PCR) typing method in which each human serotype variant of the virus produced a characteristic fragment size, which were thus readily identifiable on agarose gels. The PCR typing method was applied to all known 6 human serotypes (1, 2, 3, 4, 8, and 9); an absolute correlation was found between molecular and traditional serologic rotavirus typing methods.