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通过聚合酶链反应(PCR)和六邻体基因部分区域测序对人腺病毒进行分子分型。

Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene.

作者信息

Lu X, Erdman D D

机构信息

Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.

出版信息

Arch Virol. 2006 Aug;151(8):1587-602. doi: 10.1007/s00705-005-0722-7. Epub 2006 Feb 26.

DOI:10.1007/s00705-005-0722-7
PMID:16502282
Abstract

Human adenoviruses (Ads) are responsible for a substantial disease burden. Type-specific identification of Ads can help guide therapeutic and disease prevention strategies and aid epidemiological investigations. Immunotyping of Ads by serum neutralization (SN) is laborious and time consuming and depends upon type-specific antisera that are in short supply. A rapid molecular typing assay based on polymerase chain reaction (PCR) amplification and sequencing of Ad hexon gene hyper-variable regions 1-6 (HVR(1-6)) known to contain type-specific epitopes was evaluated as an alternative to SN. Deduced amino acid sequences of HVR(1-6) obtained from all 51 currently recognized Ad prototype strains were well resolved, with the exception of types 15 and 29, which were identical. Of 192 temporally and geographically diverse Ad field isolates sequenced in this study, and 111 previously published sequences, all more closely matched their predicted prototype strains. Ads were also detected and correctly identified directly from 24 clinical specimens positive by culture or antigen detection. PCR and sequencing of HVR(1-6) offers a practical alternative to SN for typing most Ads and can be readily adapted for use in laboratories with molecular capabilities.

摘要

人腺病毒(Ads)造成了相当大的疾病负担。腺病毒的型特异性鉴定有助于指导治疗和疾病预防策略,并有助于流行病学调查。通过血清中和(SN)对腺病毒进行免疫分型既费力又耗时,且依赖于供应短缺的型特异性抗血清。一种基于聚合酶链反应(PCR)扩增和腺病毒六邻体基因高变区1-6(HVR(1-6))测序的快速分子分型检测方法被评估为血清中和法的替代方法,已知HVR(1-6)包含型特异性表位。从目前所有51种公认的腺病毒原型株获得的HVR(1-6)推导氨基酸序列得到了很好的解析,但15型和29型除外,它们的序列相同。在本研究中测序的192株来自不同时间和地点的腺病毒现场分离株以及111条先前发表的序列中,所有序列都与其预测的原型株更紧密匹配。还直接从24份经培养或抗原检测呈阳性的临床标本中检测并正确鉴定出了腺病毒。HVR(1-6)的PCR和测序为大多数腺病毒分型提供了一种实用的血清中和法替代方法,并且可以很容易地适用于具备分子检测能力的实验室。

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