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用于快速诊断呼吸道腺病毒感染的聚合酶链反应

Polymerase chain reaction for rapid diagnosis of respiratory adenovirus infection.

作者信息

Morris D J, Cooper R J, Barr T, Bailey A S

机构信息

Department of Pathological Sciences, University of Manchester, Manchester Royal Infirmary, UK.

出版信息

J Infect. 1996 Mar;32(2):113-7. doi: 10.1016/s0163-4453(96)91250-5.

Abstract

Endemic (type 1, 2, 5 and 6) and epidemic (type 3, 4 and 7) respiratory adenovirus infections are associated with upper respiratory tract symptoms, pharyngoconjunctival fever, and pneumonia. Improved methods of diagnosis are needed, particularly in immunocompromized patients. We examined 93 throat swabs or nasopharyngeal aspirates from patients with acute respiratory disease using virus isolation and an adenovirus-specific polymerase chain reaction (PCR) based on consensus primers H1 and H2 derived from the hexon region DNA sequences of serotypes 2 and 5. Specimens which yielded viruses other than adenovirus in cell culture (n = 23) or which were negative for infectious viruses (n = 25) were negative in the PCR. The sensitivity of DNA amplification was 76% (34/45) in comparison with virus culture, being markedly lower with subgenus B (types 3 and 7) strains than with subgenus C (type 1, 2, 5 and 6) isolates (8/16 (50%)) vs. 26/28 (93%). P = 0.004) despite the use of a low annealing temperature to maximize detection of adenoviruses belonging to subgenera other than C. Of the 11 samples falsely negative in a single-round PCR but yielding adenovirus type 1 (n = 1), type 2 (n = 1). type 3 (n = 7), type 7 (n = 1), or untyped isolates (n = 1) in cell culture, nine (82%) gave positive results after nested DNA amplification. Possible approaches to further improving the performance of adenovirus PCR with respiratory specimens are discussed.

摘要

地方性(1、2、5和6型)和流行性(3、4和7型)呼吸道腺病毒感染与上呼吸道症状、咽结膜热及肺炎有关。需要改进诊断方法,尤其是在免疫功能低下的患者中。我们使用病毒分离法以及基于源自2型和5型血清型六邻体区域DNA序列的共有引物H1和H2的腺病毒特异性聚合酶链反应(PCR),检测了93例急性呼吸道疾病患者的咽拭子或鼻咽抽吸物。在细胞培养中产生除腺病毒以外病毒的标本(n = 23)或感染性病毒检测呈阴性的标本(n = 25),PCR检测结果均为阴性。与病毒培养相比,DNA扩增的灵敏度为76%(34/45),B亚属(3型和7型)毒株的灵敏度明显低于C亚属(1、2、5和6型)分离株(8/16(50%)对26/28(93%)。P = 0.004),尽管使用了低退火温度以最大限度地检测除C亚属以外的腺病毒。在单轮PCR中呈假阴性但在细胞培养中产生1型(n = 1)、2型(n = 1)、3型(n = 7)、7型(n = 1)或未分型分离株(n = 1)腺病毒的11份样本中,9份(82%)在巢式DNA扩增后呈阳性结果。本文讨论了进一步提高呼吸道标本腺病毒PCR检测性能的可能方法。

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