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用合成寡核苷酸探针检测肠道腺病毒

Detection of enteric adenoviruses with synthetic oligonucleotide probes.

作者信息

Scott-Taylor T H, Ahluwalia G, Dawood M, Hammond G W

机构信息

Cadham Provincial Laboratory, University of Manitoba, Winnipeg, Canada.

出版信息

J Med Virol. 1993 Dec;41(4):328-37. doi: 10.1002/jmv.1890410414.

Abstract

The abilities of hybridization probes to detect all human adenovirus types and to identify enteric adenovirus types were evaluated. The efficiency of hybridization was compared to other tests currently in routine laboratory use on clinical specimens from young children with gastroenteritis. Probes were derived from various regions of the adenovirus types 2 and 41 genomes, and were evaluated by hybridization with a series of DNA quantities from 1 microgram to 10 pg of one adenovirus type from each human subgenus, lambda phage, and HEp 2 cells. The sensitivity of hybridization with the HPII probe (92.7%), containing the conserved hexon gene, compared well with EM (54.6%), culture and neutralization (45.5%), and enzyme immunoassay (61.8%). The sensitivity of detection of enteric adenovirus isolates by the cloned Bg/II D fragment probe (92.9%) and by a synthetic probe (85.7%), manufactured from type-specific sequences of the Ad41 hexon gene were comparable to Ad40/Ad41 specific enzyme immunoassay (84.6%). Hybridization was found to be a sensitive method of adenovirus detection in comparison to traditional methods of laboratory diagnosis. Synthetic oligonucleotides enable specific detection of individual enteric adenovirus types. Hybridization had additional advantages over other tests in identifying cases of infection with more than one adenovirus type and in allowing an estimate of the concentration of adenovirus in the specimen.

摘要

评估了杂交探针检测所有人类腺病毒类型以及鉴定肠道腺病毒类型的能力。将杂交效率与目前实验室常规用于检测患肠胃炎幼儿临床标本的其他检测方法进行了比较。探针源自腺病毒2型和41型基因组的不同区域,并通过与来自每个人亚属、λ噬菌体和HEp 2细胞的一种腺病毒类型的一系列从1微克到10皮克的DNA量进行杂交来评估。与含有保守六邻体基因的HPII探针杂交的灵敏度(92.7%)与电子显微镜检查(54.6%)、培养和中和试验(45.5%)以及酶免疫测定(61.8%)相比表现良好。通过克隆的Bg/II D片段探针(92.9%)和由Ad41六邻体基因的型特异性序列制成的合成探针(85.7%)检测肠道腺病毒分离株的灵敏度与Ad40/Ad41特异性酶免疫测定(84.6%)相当。与传统的实验室诊断方法相比,杂交被发现是一种检测腺病毒的灵敏方法。合成寡核苷酸能够特异性检测个别肠道腺病毒类型。在鉴定感染不止一种腺病毒类型的病例以及估计标本中腺病毒的浓度方面,杂交比其他检测方法具有额外的优势。

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