ROTAVIRUS DETECTION BY DOT BLOT HYBRIDIZATION ASSAY USING A NONRADIOACTIVE SYNTHETIC OLIGODEOXYNUCLEOTIDE PROBE

FERNANDEZ, J; SANDINO, A; YUDELEVICH, A; AVENDANO, LF; VENEGAS, A; HINRICHSEN, V; SPENCER, E

ROTAVIRUS DETECTION BY DOT BLOT HYBRIDIZATION ASSAY USING A NONRADIOACTIVE SYNTHETIC OLIGODEOXYNUCLEOTIDE PROBE

Date

1992

Authors

Abstract

A synthetic oligodeoxynucleotide of 40 nucleotides corresponding to nucleotides 33-72 of the gene coding for the viral protein VP7 of rotavirus. was used as a nucleic acid probe to develop a non-radioactive hybridization method for rotavirus detection. The probe was labelled at the 3' end with biotin-7-dATP. The sensitivity and specificity of the dot blot hybridization assay for rotavirus detection was evaluated with 303 stool specimens. The results indicate that the hybridization assay has a higher sensitivity than both PAGE and EIA. Among the rotavirus strains tested 37 different electropherotypes were found. The results suggest that rotavirus diagnosis by dot hybridization using a non-radioactive probe may become routine laboratory procedure because it is simple, highly specific and very sensitive.