通过逆转录聚合酶链反应从蜜蜂中检测急性蜜蜂麻痹病毒和黑蜂王台病毒。
Detection of acute bee paralysis virus and black queen cell virus from honeybees by reverse transcriptase pcr.
作者信息
Benjeddou M, Leat N, Allsopp M, Davison S
机构信息
Department of Microbiology, University of the Western Cape, Bellville 7535, Cape Town, South Africa.
出版信息
Appl Environ Microbiol. 2001 May;67(5):2384-7. doi: 10.1128/AEM.67.5.2384-2387.2001.
Abstract
A reverse transcriptase PCR (RT-PCR) assay was developed for the detection of acute bee paralysis virus (ABPV) and black queen cell virus (BQCV), two honeybee viruses. Complete genome sequences were used to design unique PCR primers within a 1-kb region from the 3' end of both genomes to amplify a fragment of 900 bp from ABPV and 700 bp from BQCV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing ABPV and BQCV. Sensitivities were approximately 1,600 genome equivalents of purified ABPV and 130 genome equivalents of BQCV.
摘要
开发了一种逆转录酶聚合酶链反应（RT-PCR）检测方法，用于检测两种蜜蜂病毒，即急性蜜蜂麻痹病毒（ABPV）和黑蜂王台病毒（BQCV）。利用完整的基因组序列在两个基因组3'端1 kb区域内设计独特的PCR引物，以扩增来自ABPV的900 bp片段和来自BQCV的700 bp片段。采用硫氰酸胍和硅胶膜联合方法从健康和实验室感染的蜜蜂蛹样本中提取总RNA。在一次盲测中，RT-PCR成功鉴定出含有ABPV和BQCV的样本。灵敏度分别约为1600个纯化ABPV基因组当量和130个BQCV基因组当量。
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