采用 PCR 法和常规实验室诊断工具评估疑似病毒性角膜炎患者的泪液样本中单纯疱疹病毒 1（HSV）的检测。

Evaluation of tear samples for Herpes Simplex Virus 1 (HSV) detection in suspected cases of viral keratitis using PCR assay and conventional laboratory diagnostic tools.

机构信息

Department of Ocular Microbiology, Dr R P Center for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.

出版信息

Br J Ophthalmol. 2011 Mar;95(3):415-8. doi: 10.1136/bjo.2010.191049. Epub 2010 Sep 18.

DOI:10.1136/bjo.2010.191049

PMID:20852317

Abstract

BACKGROUND

Herpes Simplex Virus (HSV) keratitis is a leading cause of corneal blindness. Definitive laboratory diagnosis is essential for timely management. Collection of corneal scrapings in patients with advanced epithelial keratitis and corneal thinning poses perforation risks; tear fluid is a feasible and convenient alternative but has not been widely evaluated for HSV detection.

METHODS

Tear fluid alone (229) or along with corneal scrapings (153) from patients of suspected herpetic keratitis was tested for HSV-1 antigen by indirect immunofluorescence assay, virus isolation in Hep 2 cells and PCR to amplify the 111bp region of the thymidine kinase (tk) coding gene and the 144bp region from the DNA polymerase coding gene of HSV.

RESULTS

HSV 1 antigen was detected in 31/229 (13.53%) tear specimen and 35/153 (22.87%) corneal scrapings in immunofluorescence assay; virus was isolated from 12/229 (5.2%) tear and 17/153 (11.11%) corneal scrapings, and PCR was positive for both the genes in 32/229 (13.97%) tear specimen and 56/153 (36.66%) corneal scrapings.

CONCLUSION

Corneal scrapings yielded a significantly better HSV positivity than tears in both the PCR assay (p<0.0005) and immunofluorescence assay. PCR was much more sensitive than immunofluorescence and virus isolation. However, tears should be tested for definitive laboratory diagnosis of HSV infection whenever corneal scraping collection is not possible.

摘要

背景

单纯疱疹病毒（HSV）角膜炎是导致角膜盲的主要原因。明确的实验室诊断对于及时治疗至关重要。对于进展性上皮性角膜炎和角膜变薄的患者，采集角膜刮片有穿孔风险；而泪液是一种可行且方便的替代方法，但尚未广泛评估其在 HSV 检测中的应用。

方法

对疑似单纯疱疹性角膜炎患者的单纯泪液（229 例）或同时采集的角膜刮片（153 例），采用间接免疫荧光法检测 HSV-1 抗原，在 Hep 2 细胞中进行病毒分离，并通过聚合酶链反应（PCR）扩增 HSV 的胸苷激酶（tk）编码基因的 111bp 区域和 DNA 聚合酶编码基因的 144bp 区域。

结果

免疫荧光法检测到 31/229（13.53%）份泪液标本和 35/153（22.87%）份角膜刮片中存在 HSV 1 抗原；从 12/229（5.2%）份泪液和 17/153（11.11%）份角膜刮片中分离出病毒，PCR 对这两个基因均呈阳性的分别有 32/229（13.97%）份泪液标本和 56/153（36.66%）份角膜刮片。