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

机译:通过聚合酶链反应检测人脑石蜡切片中人巨细胞病毒DNA的发生和假阴性结果的发生。

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摘要

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.
机译:通过巢式聚合酶链反应(nPCR)检查了6例免疫细胞化学确诊的人巨细胞病毒性脑炎(HCMVE)患者和11例对照石蜡包埋的尸检材料。在4例HCMVE中,HCMV基因组的特征性183个碱基对(bp)片段很容易扩增。在2例HCMVE病例中,由于DNA提取效率低下或DNA降解最可能仅通过PCR偶尔显示出病毒DNA。所有对照病例均为阴性。 nPCR提供了一种检测常规处理的活检和尸检材料中HCMV DNA的特定方法,可用于档案组织中以诊断感染。但是,样品的固定和DNA提取是关键步骤,如果将PCR用于检测HCMV,则需要仔细控制,以避免假阴性结果。

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