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通过酶免疫测定法对临床样本中的单纯疱疹病毒进行快速检测、培养扩增及分型

Rapid detection, culture-amplification and typing of herpes simplex viruses by enzyme immunoassay in clinical samples.

作者信息

Kok T, Mickan L, Schepetiuk S

机构信息

Infectious Diseases Laboratory, Institute of Medical and Veterinary Science, Adelaide, South Australia.

出版信息

Clin Diagn Virol. 1998 May 1;10(1):67-74. doi: 10.1016/s0928-0197(98)00003-8.

Abstract

BACKGROUND

The laboratory diagnosis of herpes simplex infection may require rapid (direct) tests, as well as cell cultures, for detection of the virus in clinical samples. The quantity of virus present in clinical samples is variable and this may depend on the period from onset of rash. In addition, not all patients may show obvious symptoms with this infection. The successful culture of herpes simplex virus requires prompt transportation after collection of the specimen as the virus is easily inactivated. Hence, rapid and culture tests would enable detection of non-viable and viable viruses.

STUDY DESIGN

We describe the rapid detection of HSV by EIA directly in various clinical samples using commercially available polyclonal sera. In addition specimens were inoculated in microwell cell cultures and 4 days post inoculation the culture fluids were tested for HSV and subtyped by a similar EIA (culture amplified EIA).

RESULTS

The direct EIA showed an endpoint detection of 100 TCID50/ml, sensitivity of 92% (all specimen types) and specificity of 100%. The direct EIA sensitivity was 97% in non-genital specimens and 88% in genital specimens. The culture amplified EIA showed a sensitivity of 95% compared to all confirmed HSV positive samples.

CONCLUSIONS

The results of the HSV rapid tests were available within 24 h from receipt of specimens. Specimens which were culture negative/direct EIA positive were confirmed by blocking antisera. Culture positive specimens which were direct EIA negative were confirmed by subtyping of the virus.

摘要

背景

单纯疱疹感染的实验室诊断可能需要快速(直接)检测以及细胞培养,以在临床样本中检测病毒。临床样本中存在的病毒量各不相同,这可能取决于出疹后的时间。此外,并非所有患者感染此病毒时都会出现明显症状。单纯疱疹病毒的成功培养需要在采集标本后迅速送检,因为该病毒很容易失活。因此,快速检测和培养检测能够检测出无活性和有活性的病毒。

研究设计

我们描述了使用市售多克隆血清通过酶免疫测定法(EIA)直接在各种临床样本中快速检测单纯疱疹病毒(HSV)。此外,将标本接种到微孔细胞培养物中,接种后4天检测培养液中的HSV,并通过类似的EIA(培养扩增EIA)进行亚型鉴定。

结果

直接EIA的终点检测为100 TCID50/ml,敏感性为92%(所有样本类型),特异性为100%。直接EIA在非生殖器标本中的敏感性为97%,在生殖器标本中的敏感性为88%。与所有确诊的HSV阳性样本相比,培养扩增EIA的敏感性为95%。

结论

HSV快速检测结果可在收到标本后24小时内获得。培养阴性/直接EIA阳性的标本通过封闭抗血清进行确认。培养阳性而直接EIA阴性的标本通过病毒亚型鉴定进行确认。

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