Experience with serotyping rotavirus strains by reverse transcription and two-step polymerase chain reaction with generic and type-specific primers.
Six VP7 serotypes or G types (G1-G4, G8 and G9) occur in group A human rotaviruses. Gouvea et al. recently reported a novel G-typing method based on reverse transcription (RT)-polymerase chain reaction (PCR) amplification of the VP7 gene with type-specific primers [Gouvea, V. et al. (1990). Journal of Clinical Microbiology 28, 276-82]. When we followed their protocol, 40 (89%) of 45 faecal rotavirus specimens were typed into G1-G4 and G9. The five specimens that were untypeable by the RT-PCR method contained three G2 and two G4 rotavirus specimens which were identified by an ELISA using G type-specific monoclonal antibodies. On the other hand, the RT-PCR typing assay was able to determine the G type of the seven isolates that were untypeable by the ELISA. Of 33 faecal rotaviruses that were typed by both assays, 100% agreement of the result was observed. In addition, when applied to some animal rotaviruses, the RT-PCR method identified G3 feline and canine rotavirus strains. We conclude that further refinement of the RT-PCR assay is desirable in order to more readily type G2 and G4 strains, although this assay displayed an applicability to epidemiologic studies comparable with ELISAs.
Nakagomi
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Department of Laboratory Medicine, Akita University School of Medicine, Japan.
Oyamada
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Nakagomi
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D004279:DNA, Viral / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005243:Feces / Q000382:microbiology; D015684:Gene Products, pol; D006801:Humans; D007564:Japan / Q000453:epidemiology; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase; D012401:Rotavirus / Q000145:classification*; D012400:Rotavirus Infections / Q000175:diagnosis* / Q000453:epidemiology; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods*; D014158:Transcription, Genetic
D004279:DNA, Viral; D015684:Gene Products, pol; D012194:RNA-Directed DNA Polymerase