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Typing of human group A rotavirus with alkaline phosphatase‐labeled oligonucleotide probes
Author(s) -
Sethabutr Orntipa,
Hanchalay Suvath,
Lexomboon Udom,
Bishop Ruth F.,
Holmes Ian H.,
Echeverria Peter
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.1890370308
Subject(s) - typing , oligonucleotide , rotavirus , virology , alkaline phosphatase , reoviridae , biology , microbiology and biotechnology , virus , enzyme , genetics , gene , biochemistry
Abstract Rotavirus (RV) in stools of children <1 year of age with diarrhea in Bangkok in 1989 were serotyped by monoclonal enzyme immunoassay (MEIA). RNA extracted from these specimens was tested for hybridization with alkaline phosphatase (AP) and 32 P‐labeled oligonucleotides constructed from the nucleotide sequences of VP7 of human G types 1 (HuG1Ac), 2 (HuG2Ac), 3 (HuG3Ac), and 4 (HuG4Ac). Of 148 specimens that contained RV, 72% (106/148) hybridized with RV G type specific AP‐labeled oligonucleotides compared to 47% (70/148) that were serotyped by MEIA ( P < 0.001). Of 68 specimens that contained only one VP7 serotype (G‐type), as identified by MEIA, 94% (16/17) of G1, 90% (27130) of G2, 57% (4/7) of G3, and 36% (5/14) of G4 RV hybridized with the AP‐labeled HuG1Ac, HuG2Ac, HuG3Ac, and HuG4Ac oligonucleotides, respectively. The probes for G1, 2,3, and 4 RV were specific for each G type. The results of hybridizing specimens with 32 P‐ and AP‐labeled oligonucleotides were similar. After transcription and amplification of cDNA of gene 9, AP‐labeled RV G type specific oligonucleotides hybridized with 90% (134/148) of RV specimens. The high sensitivity of these nonimmunological techniques could be of value in identifying G types of RV during vaccine trials. © 1992 Wiley‐Liss, Inc.