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根据临床表现进行腺病毒感染诊断的实验室方法。

Laboratory approaches to the diagnosis of adenovirus infection depending on clinical manifestations.

作者信息

Terletskaia-Ladwig E, Leinmüller M, Schneider F, Meier S, Enders M

机构信息

Labor Enders und Partner, Institut für Virologie, Infektiologie und Epidemiologie e.V., Stuttgart, Germany.

出版信息

Infection. 2007 Dec;35(6):438-43. doi: 10.1007/s15010-007-6340-4. Epub 2007 Oct 9.

Abstract

BACKGROUND

While commercial enzyme immunoassays (EIA) intended for the detection of adenovirus in fecal specimens are widely used, there are no rapid, convenient, and sensitive commercial tests available for the detection of adenoviruses in respiratory and conjunctival specimens. The applicability of EIA for the detection of adenovirus in stool and throat samples was investigated. One-day rapid culture assay (RCA) for the detection of adenovirus in respiratory and conjunctival specimens was developed and evaluated.

PATIENTS AND METHODS

Stool samples from patients with gastroenteritis were tested by adenovirus EIA and by cell culture using human embryonic lung cells (HEL) and Graham 293 cells. Blood and stool samples from two BMT patients were also tested for adenovirus by PCR for at least 6 months. Throat specimens from patients with respiratory infections and conjunctival specimens were used for the evaluation of 1-day RCA compared with conventional adenovirus isolation in Graham 293 cells.

RESULTS

A total of 3,860 stool samples were tested by EIA Ridascreen, 8,169 by Novitec, and 2,218 by ProSpectT yielding 135 (3.5%), 308 (3.7%), and 77 (3.5%) positive results, respectively. From 305 Ridascreen- and 340 Novitec-negative stool samples, adenoviruses were isolated in three (0.9%) and eight (2.4%) cases, respectively, including two patients undergoing BMT. Multiple sequential stool samples from one BMT patient were repeatedly negative by EIA, but positive by PCR and cell culture. Graham 293 cells were better suited for isolation of adenovirus than HEL. EIA proved unreliable for detecting adenovirus in throat swabs. The sensitivity and specificity of RCA in throat swabs were 90% (37/41) and 100% (64/64), respectively, and 76% (16/21) and 100% (132/132) in conjunctival specimens, respectively.

CONCLUSIONS

Generally, EIA is sufficiently sensitive for the diagnosis of adenovirus-associated diarrhea. However, it may not be sensitive enough to detect adenovirus in immunocompromised patients undergoing BMT and shedding very few viral particles in stools. Thus, in such cases, a more sensitive assay, such as PCR, is recommended. Furthermore, EIA is not sufficiently sensitive for the reliable detection of adenoviruses in throat swabs. One-day RCA may be useful for the detection of adenoviruses in respiratory and conjunctival specimens.

摘要

背景

虽然用于检测粪便标本中腺病毒的商业酶免疫测定法(EIA)被广泛使用,但目前尚无快速、便捷且灵敏的商业检测方法可用于检测呼吸道和结膜标本中的腺病毒。我们研究了EIA在检测粪便和咽喉样本中腺病毒的适用性。同时开发并评估了一种用于检测呼吸道和结膜标本中腺病毒的一日快速培养测定法(RCA)。

患者与方法

对肠胃炎患者的粪便样本采用腺病毒EIA以及使用人胚肺细胞(HEL)和格雷厄姆293细胞进行细胞培养检测。还对两名骨髓移植(BMT)患者的血液和粪便样本进行了至少6个月的腺病毒PCR检测。将呼吸道感染患者的咽喉标本和结膜标本用于评估一日RCA,并与在格雷厄姆293细胞中进行的传统腺病毒分离方法作比较。

结果

使用Ridascreen EIA检测了3860份粪便样本,Novitec检测了8169份,ProSpectT检测了2218份,阳性结果分别为135份(3.5%)、308份(3.7%)和77份(3.5%)。在305份Ridascreen检测阴性和340份Novitec检测阴性的粪便样本中,分别有3例(0.9%)和8例(2.4%)分离出腺病毒,其中包括两名接受BMT的患者。一名BMT患者的多个连续粪便样本经EIA检测反复呈阴性,但PCR和细胞培养检测呈阳性。格雷厄姆293细胞比HEL更适合分离腺病毒。EIA被证明在检测咽喉拭子中的腺病毒时不可靠。RCA在咽喉拭子中的敏感性和特异性分别为90%(37/41)和100%(64/64),在结膜标本中分别为76%(16/21)和100%(132/132)。

结论

一般来说,EIA对腺病毒相关腹泻的诊断具有足够的敏感性。然而,对于接受BMT且粪便中病毒颗粒极少的免疫功能低下患者,它可能不够灵敏以检测到腺病毒。因此,在这种情况下,建议采用更灵敏的检测方法,如PCR。此外,EIA对于可靠检测咽喉拭子中的腺病毒不够灵敏。一日RCA可能有助于检测呼吸道和结膜标本中的腺病毒。

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