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聚合酶链反应与标准Southern印迹法在检测各种活检标本中EB病毒DNA的比较。

Comparison of polymerase chain reaction and standard Southern blotting for the detection of Epstein-Barr virus DNA in various biopsy specimens.

作者信息

Pedneault L, Katz B Z

机构信息

Department of Medical Microbiology, Université de Montréal, Québec, Canada.

出版信息

J Med Virol. 1993 Jan;39(1):33-43. doi: 10.1002/jmv.1890390108.

Abstract

The sensitivity of the polymerase chain reaction (PCR) assay was compared to that of standard Southern blotting (SB) hybridization for detecting the presence of Epstein-Barr virus (EBV) genomes in biopsy samples from 43 patients with a variety of lymphoproliferative disorders. Two pairs of oligonucleotide primers from the first BamHI M and R leftward reading frames (BMLF1 and BRLF1) of EBV were chosen to amplify DNA. The resulting PCR products were analyzed by gel electrophoresis, transfer and hybridization. Restriction enzyme digestion was used to confirm the specificity of the amplified fragment. EBV DNA was found in 38 of 43 patients, as compared with 9 of 43 patients with the Southern technique. No amplified product was detected with other viruses from the Herpes family, nor with human genomic DNA from healthy adults using the same two sets of primers. These results indicate that EBV can be detected in a greater number of lymphoproliferative lesions than previously appreciated. The implications of these findings are discussed.

摘要

将聚合酶链反应(PCR)检测的灵敏度与标准Southern印迹(SB)杂交法进行比较,以检测43例患有各种淋巴增生性疾病患者活检样本中Epstein-Barr病毒(EBV)基因组的存在情况。选择来自EBV的第一个BamHI M和R左向读框(BMLF1和BRLF1)的两对寡核苷酸引物来扩增DNA。对所得的PCR产物进行凝胶电泳、转移和杂交分析。使用限制性内切酶消化来确认扩增片段的特异性。Southern印迹技术检测到43例患者中有9例存在EBV DNA,而PCR检测到43例患者中有38例存在EBV DNA。使用相同的两组引物,未检测到来自疱疹病毒家族的其他病毒以及健康成年人的人类基因组DNA的扩增产物。这些结果表明,与之前的认识相比,在更多的淋巴增生性病变中可以检测到EBV。讨论了这些发现的意义。

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