Coulson B S, Gentsch J R, Das B K, Bhan M K, Glass R I
Department of Microbiology and Immunology, the University of Melbourne, Parkville 3052, Victoria, Australia.
J Clin Microbiol. 1999 Oct;37(10):3187-93. doi: 10.1128/JCM.37.10.3187-3193.1999.
While only four globally important rotavirus G serotypes (1 to 4) have been documented, many studies suggest that serotype G9 viruses may be widely distributed and more important than previously recognized. We have evaluated 10 serotype G9 rotavirus-neutralizing monoclonal antibodies (MAbs) directed to VP7, which bound by direct enzyme immunoassay (EIA) to P1A[8], G9 rotaviruses F45, WI61, and AU32, for their ability to recognize the New Delhi G9 rotavirus 116E. Only one MAb (MAb F45:1) bound to P[11], G9 virus 116E to a high titer by EIA. This MAb was incorporated into an indirect EIA for G serotyping, which was validated with prototype cultivable human rotaviruses of G types 1 to 4 and 9. The EIA was compared with genotyping by reverse transcriptase PCR (RT-PCR) under code for the determination of the G types of rotaviruses obtained from neonates in New Delhi, India. The sensitivities of RT-PCR and EIA (after two additional freeze-thaw cycles) for the typing of G9 rotaviruses were 91 and 86%, respectively, for 24 culture-adapted rotavirus strains. The untypeable culture-adapted rotavirus samples also were unreactive with VP7 group antigen-reactive MAb 60. After two additional freeze-thaw cycles, only 26 of 42 (62%) of stools containing rotavirus typed as G9 by RT-PCR were positive for G9 rotavirus by EIA. Stools containing rotavirus untypeable by EIA contained significantly less MAb 60-reactive VP7 antigen (P = 0. 0001) than the stools containing typeable rotavirus. Thus, RT-PCR genotyping was the more sensitive method for determination of G9 type, but a serotype was readily determined in rotavirus samples containing MAb 60-reactive VP7 antigen by an EIA that incorporates MAb F45:1.
虽然全球仅记录了4种重要的轮状病毒G血清型（1至4型），但许多研究表明，G9血清型病毒可能分布广泛，且比以前认为的更为重要。我们评估了10种针对VP7的G9血清型轮状病毒中和单克隆抗体（MAb），这些抗体通过直接酶免疫测定（EIA）与P1A[8]、G9轮状病毒F45、WI61和AU32结合，以检测它们识别新德里G9轮状病毒116E的能力。通过EIA，只有一种单克隆抗体（MAb F45:1）能与P[11]、G9病毒116E高滴度结合。该单克隆抗体被用于建立一种间接EIA进行G血清型分型，并通过G1至4型和9型的原型可培养人轮状病毒进行验证。在对从印度新德里新生儿中获得的轮状病毒进行G血清型测定时，将该EIA与逆转录酶PCR（RT-PCR）基因分型法进行了盲法比较。对于24株适应细胞培养的轮状病毒株，RT-PCR和EIA（经过另外两个冻融循环）对G9轮状病毒分型的敏感性分别为91%和86%。无法分型的适应细胞培养的轮状病毒样本与VP7组抗原反应性单克隆抗体60也无反应。经过另外两个冻融循环后，通过RT-PCR分型为G9的42份含轮状病毒粪便样本中，只有26份（62%）通过EIA检测为G9轮状病毒阳性。EIA无法分型的含轮状病毒粪便样本中，MAb 60反应性VP7抗原的含量明显低于可分型轮状病毒的粪便样本（P = 0.0001）。因此，RT-PCR基因分型是确定G9型的更敏感方法，但对于含有MAb 60反应性VP7抗原的轮状病毒样本，通过包含MAb F45:1的EIA可以很容易地确定其血清型。
