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Respiratory syncytial virus detection by dot blot hybridization with a nonradioactive synthetic oligo deoxynucleotide probe
Author(s) -
Hernández Oscar,
Fernandez Jorge,
Valenzuela Sofia,
Sandino Ana Maria,
Pizarro Jaqueline,
Vasquez Monica,
Yudelevich Arturo,
Spencer Eugenio
Publication year - 1992
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890370303
Subject(s) - dot blot , virology , immunofluorescence , virus , microbiology and biotechnology , biology , hybridization probe , southern blot , paramyxoviridae , rotavirus , molecular probe , western blot , dna , antibody , gene , viral disease , immunology , biochemistry
Abstract A synthetic oligodeoxynucleotide corresponding to a region of the nucleocapside gene (N) of respiratory syncytial virus (RSV), was used as a DNA probe to develop a nonradioactive hybridization assay for the detection of RSV. The probe was labeled by incorporation of biotin‐7‐dATP to the 3′ end by a reaction catalyzed by terminal deoxynucleotydil transferase. The dot blot hybridization assay was found to be specific for RSV when tested against RSV isolates (subgroups A and B) obtained from cell cultures and isolates of adenovirus, reovirus, rotavirus, and pararotavirus. The assay detected both RSV subgroups (A and B) without significant differences. The dot blot hybridization assay using the nonradioactive probe led to similar results to indirect immunofluorescence (IFI) when tested against a panel of 64 clinical samples from nasopharyngeal secretions of infants with clinical symptoms of respiratory disease. This assay may provide the basis for a rapid, simple, and inexpensive method for routine RSV diagnosis. © 1992 Wiley‐Liss, Inc.