Rotavirus detection by dot blot hybridization assay using a non-radioactive synthetic oligodeoxynucleotide probe.
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.
Fernández
J
J
Unidad de Virología INTA, Universidad de Chile, Santiago.
Sandino
A
A
Yudelevich
A
A
Avendaño
L F
LF
Venegas
A
A
Hinrichsen
V
V
Spencer
E
E
D001483:Base Sequence; D003967:Diarrhea / Q000175:diagnosis; D004591:Electrophoresis, Polyacrylamide Gel; D005243:Feces / Q000382:microbiology; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000737:chemistry*; D011237:Predictive Value of Tests; D012330:RNA, Double-Stranded / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis* / Q000737:chemistry; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis*
D015345:Oligonucleotide Probes; D012330:RNA, Double-Stranded; D012367:RNA, Viral