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1993 - 1994年期间住院的印度儿童不可分型粪便标本中轮状病毒AU32样G9毒株的证据。

Evidence of rotavirus AU32 like G9 strains from nontypeable fecal specimens of Indian children hospitalized during 1993-1994.

作者信息

Awachat Pinki S, Kelkar Shobhana D

机构信息

National Institute of Virology, Pune, India.

出版信息

J Med Virol. 2004 Dec;74(4):656-61. doi: 10.1002/jmv.20227.

Abstract

Serotyping of 432 rotavirus positive fecal specimens collected from hospitalized children during 1990-1997 was carried out at National Institute of Virology (NIV), Pune, India, using monoclonal antibodies (MAbs) directed against VP7 determinant of serotypes G1-G4, G6, G8, and G10. However, significant number of specimens, that is, 47.92% remained nontypeable. The aim of the present study was to culture adapt the nontypeable specimens and to characterize them further. Since the fecal specimens were not tested by MAb to G9 serotype, which has emerged as an important serotype infecting humans recently, presence of G9 serotype was expected in nontypeable specimens. Therefore, we selected specimens from those children, who showed higher neutralizing antibody (NAb) titer in their convalescent serum samples to G9 serotype than their mothers. Out of six isolates having long electropherotype, five isolates showed subgroup II, and one showed subgroup I, II. The isolates were confirmed as G9 by MAb based ELISA, neutralization assay, and PCR. The G9 specific nested PCR products of four isolates showed 96-99% identities to AU32 G9 strain reported from Japan. P type of four isolates was determined as P8. Besides isolates, four additional nontypeable fecal specimens were confirmed as G9 by MAb based ELISA. Thus, 10 (28.57%) out of 35 nontypeable specimens were identified as rotavirus serotype G9. The results indicate that serotype G9 may represent significant proportion of specimens, which were previously nontypeable.

摘要

1990年至1997年期间,从印度浦那国立病毒研究所(NIV)住院儿童中收集了432份轮状病毒阳性粪便标本,使用针对G1 - G4、G6、G8和G10血清型VP7决定簇的单克隆抗体(MAb)进行血清分型。然而,相当数量的标本,即47.92%,仍无法分型。本研究的目的是对无法分型的标本进行培养适应性处理并进一步鉴定。由于粪便标本未用针对G9血清型的单克隆抗体进行检测,而G9血清型最近已成为感染人类的重要血清型,因此预计无法分型的标本中存在G9血清型。因此,我们从那些恢复期血清样本中对G9血清型的中和抗体(NAb)滴度高于其母亲的儿童中选择标本。在六个具有长电泳型的分离株中,五个分离株显示为II亚组,一个显示为I、II亚组。通过基于单克隆抗体的ELISA、中和试验和PCR确认分离株为G9。四个分离株的G9特异性巢式PCR产物与日本报道的AU32 G9株具有96 - 99%的同一性。四个分离株的P型被确定为P8。除了分离株外,另外四份无法分型的粪便标本通过基于单克隆抗体的ELISA被确认为G9。因此,35份无法分型的标本中有10份(28.57%)被鉴定为轮状病毒G9血清型。结果表明,G9血清型可能占以前无法分型标本的很大比例。

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