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通过聚合酶链反应检测人脑海绵状血管病石蜡切片中的人巨细胞病毒DNA及假阴性结果的发生情况

Detection of human cytomegalovirus DNA in paraffin sections of human brain by polymerase chain reaction and the occurrence of false negative results.

作者信息

Gass P, Kiessling M, Schäfer P, Mester C, Schmitt H P, Kühn J E

机构信息

University of Heidelberg, Institute of Neuropathology, Germany.

出版信息

J Neurol Neurosurg Psychiatry. 1993 Feb;56(2):211-4. doi: 10.1136/jnnp.56.2.211.

Abstract

Paraffin-embedded necropsy material from 6 patients with human cytomegalovirus encephalitis (HCMVE) corroborated by immunocytochemistry and 11 control cases were examined for the presence of human cytomegalovirus (HCMV) DNA by a nested polymerase chain reaction (nPCR). A characteristic 183 base pair (bp) fragment of the HCMV genome could readily be amplified in 4 cases of HCMVE. In 2 cases of HCMVE, viral DNA could be demonstrated only sporadically by PCR, due most likely to inefficient DNA extraction or DNA degradation. All control cases remained negative. The nPCR provides a specific method for detecting HCMV DNA in routinely processed biopsy and necropsy material and may be used in archival tissues for the diagnosis of infection. Fixation of samples and DNA extraction are, however, crucial steps and require careful control if PCR is used for detection of HCMV, to avoid false negative results.

摘要

通过免疫细胞化学证实的6例人类巨细胞病毒脑炎(HCMVE)患者的石蜡包埋尸检材料和11例对照病例,采用巢式聚合酶链反应(nPCR)检测人巨细胞病毒(HCMV)DNA的存在情况。HCMV基因组的一个特征性183碱基对(bp)片段在4例HCMVE中很容易被扩增出来。在2例HCMVE中,病毒DNA仅通过PCR偶尔被检测到,最可能的原因是DNA提取效率低下或DNA降解。所有对照病例均为阴性。nPCR为在常规处理的活检和尸检材料中检测HCMV DNA提供了一种特异性方法,可用于存档组织中感染的诊断。然而,如果使用PCR检测HCMV,样本固定和DNA提取是关键步骤,需要仔细控制以避免假阴性结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a3dc/1014827/947d94ab5dd7/jnnpsyc00475-0093-a.jpg

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