免疫荧光和PCR检测存档石蜡包埋脑组织狂犬病的方法评价。

K Kulonen, M Fekadu, S Whitfield, C K Warner
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引用次数: 18

摘要

比较了直接免疫荧光和PCR两种检测方法对石蜡包埋头颅标本的敏感性。这些材料包括1988年芬兰狂犬病爆发期间获得的23份样本,以及1985年从芬兰死于狂犬病的一名蝙蝠研究人员身上分离出的一份样本。这些结果与对新鲜组织进行的原始诊断进行了比较。免疫荧光法检出率为100%(12/12)。设计用于检测狂犬病核蛋白基因5'端附近139 bp靶点的PCR检测也在新鲜组织标本中检测到100%(12/12)的阳性样本。一项旨在检测304-bp靶标的PCR检测跨越了第一项检测的139-bp靶标,仅检测到67%(8/12)的原始病例。无假阳性记录。抗原的免疫荧光检测和核蛋白基因短区域的PCR检测都可用于确定固定石蜡包埋(档案)材料的狂犬病状态。
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An evaluation of immunofluorescence and PCR methods for detection of rabies in archival Carnoy-fixed, paraffin-embedded brain tissue.

Direct immunofluorescence and PCR detection methods were compared for sensitivity in evaluating the rabies status of archival specimens of Carnoy-fixed, paraffin-embedded brain tissue. The material consisted of 23 samples obtained during a rabies outbreak in Finland in 1988, and one sample isolated from a bat researcher who died of rabies in Finland in 1985. These results were compared with the original diagnoses performed on the fresh tissues. The immunofluorescence assay detected 100% (12/12) of the rabies-positive archival cases. A PCR assay designed to detect a 139-bp target near the 5' end of the rabies nucleoprotein gene also detected 100% (12/12) of the samples identified as positive in the fresh tissue specimens. A PCR assay designed to detect a 304-bp target spanning the 139-bp target of the first assay detected only 67% (8/12) of the original cases. No false positives were recorded. Both immunofluorescence detection of antigen and PCR detection of a short region of the nucleoprotein gene are useful in determining the rabies status of fixed, paraffin embedded (archival) material.

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