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Rotavirus detection by dot blot hybridization assay using a non-radioactive synthetic oligodeoxynucleotide probe

Published online by Cambridge University Press:  15 May 2009

J. Fernández ,
A. Sandino ,
A. Yudelevich ,
L. F. Avendaño ,
A. Venegas ,
V. Hinrichsen  and
E. Spencer
J. Fernández
Affiliation:
Unidad de Virologia INTA, Universidad de Chile, Macul 5540, Casilla 138–11, Santiago, Chile.
A. Sandino
Affiliation:
Unidad de Virologia INTA, Universidad de Chile, Macul 5540, Casilla 138–11, Santiago, Chile.
A. Yudelevich
Affiliation:
Departamento de Biologia Celular y Molecular, Facultad de Ciencias Biologicas, Pontificia Universidad Catolica de Chile
L. F. Avendaño
Affiliation:
Unidad de Virologia INTA, Universidad de Chile, Macul 5540, Casilla 138–11, Santiago, Chile.
A. Venegas
Affiliation:
Departamento de Biologia Celular y Molecular, Facultad de Ciencias Biologicas, Pontificia Universidad Catolica de Chile
V. Hinrichsen
Affiliation:
Departamento de Biologia Celular y Molecular, Facultad de Ciencias Biologicas, Pontificia Universidad Catolica de Chile
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Summary

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A synthetic oligodeoxynucletide 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-radiactive 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.

Type
Research Article
Information
Epidemiology & Infection , Volume 108 , Issue 1 , February 1992 , pp. 175 - 184
Copyright
Copyright © Cambridge University Press 1992

References

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