Typing of human group A rotavirus with alkaline phosphatase-labeled oligonucleotide probes

✍ Scribed by Orntipa Sethabutr; Suvath Hanchalay; Udom Lexomboon; Ruth F. Bishop; Ian H. Holmes; Peter Echeverria

• Book ID: 102910239
• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 564 KB
• Volume: 37
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/jmv.1890370308

✦ Synopsis

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.