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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.

DOI:10.1002/jmv.1890410414
PMID:8106869
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7166767/
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。

相似文献

1
Detection of enteric adenoviruses with synthetic oligonucleotide probes.用合成寡核苷酸探针检测肠道腺病毒
J Med Virol. 1993 Dec;41(4):328-37. doi: 10.1002/jmv.1890410414.
2
DNA hybridization for diagnosis of enteric adenovirus infection from directly spotted human fecal specimens.通过直接点样的人类粪便标本进行DNA杂交以诊断肠道腺病毒感染。
J Clin Microbiol. 1987 Oct;25(10):1881-5. doi: 10.1128/jcm.25.10.1881-1885.1987.
3
Specific detection and typing of adenovirus types 40 and 41 in stool specimens by dot-blot hybridization.通过斑点杂交法对粪便标本中的40型和41型腺病毒进行特异性检测和分型。
J Clin Microbiol. 1985 Dec;22(6):934-9. doi: 10.1128/jcm.22.6.934-939.1985.
4
Isolation and propagation of enteric adenoviruses in HEp-2 cells.肠道腺病毒在人喉表皮样癌细胞(HEp-2细胞)中的分离与增殖
J Clin Microbiol. 1988 Aug;26(8):1445-7. doi: 10.1128/jcm.26.8.1445-1447.1988.
5
Detection of adenoviruses in stool specimens by nucleic acid spot hybridization.通过核酸斑点杂交法检测粪便标本中的腺病毒。
J Med Virol. 1985 Jul;16(3):213-8. doi: 10.1002/jmv.1890160302.
6
Prevalent enteric adenovirus variant not detected by commercial monoclonal antibody enzyme immunoassay.
J Clin Microbiol. 1990 Dec;28(12):2797-801. doi: 10.1128/jcm.28.12.2797-2801.1990.
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Direct detection and differentiation of fastidious and nonfastidious adenoviruses in stools by using a specific nonradioactive probe.使用特异性非放射性探针直接检测和区分粪便中苛养型和非苛养型腺病毒。
J Clin Microbiol. 1986 Nov;24(5):785-9. doi: 10.1128/jcm.24.5.785-789.1986.
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Comparison of detection methods for adenovirus from enteric clinical specimens.肠道临床标本中腺病毒检测方法的比较
Diagn Microbiol Infect Dis. 1994 Mar;18(3):161-6. doi: 10.1016/0732-8893(94)90086-8.
9
Detection, typing, and subtyping of enteric adenoviruses 40 and 41 from fecal samples and observation of changing incidences of infections with these types and subtypes.从粪便样本中检测、分型和进一步细分肠道腺病毒40型和41型,并观察这些型别和亚型感染发生率的变化。
J Clin Microbiol. 1993 Jun;31(6):1562-9. doi: 10.1128/jcm.31.6.1562-1569.1993.
10
Laboratory identification of adenoviruses associated with gastroenteritis in Canada from 1983 to 1986.1983年至1986年加拿大与肠胃炎相关的腺病毒的实验室鉴定
J Clin Microbiol. 1990 Jul;28(7):1525-9. doi: 10.1128/jcm.28.7.1525-1529.1990.

本文引用的文献

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