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首页> 外文期刊>Journal of Clinical Microbiology >Use of multiple PCR primer sets for optimal detection of human papillomavirus.
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Use of multiple PCR primer sets for optimal detection of human papillomavirus.

机译：使用多种PCR引物组对人乳头瘤病毒进行最佳检测。

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Using multiple PCR primer sets, we tried to optimize the detection of human papillomavirus (HPV) in DNA samples isolated from 361 frozen biopsy specimens from patients with invasive cervical carcinomas. The HPVs detected were placed into three distinct groups, including group I/Inex at Telelab (Skien, Norway) and group Ineg and group II at the Norwegian Radium Hospital (Oslo, Norway). The consensus primer sets were Oli-1b-oli-2i, My09-My11, Gp5-Gp6, and Gp(5+)-Gp6+ from the HPV L1 gene and CpI-CpIIG from the E1 gene. Using these consensus primers together with the type-specific primers from E6-E7, we found that 355 patients (98%) were HPV positive. Type-specific primers for HPV types 11, 16, 18, 31, 33, and 35 detected more HPV-infected patients than the most sensitive consensus primer set, while the three consensus primer sets My, Gp/Gp+, and Cp together detected more HPV-positive patients than the type-specific primers. Testing of sensitivity of the PCR with SiHa cells serially diluted in lymphocytes (HPV-negative cells) indicated a detection limit of 6,300 HPV type 16 DNA copies with consensus primers (My, Gp+, and Cp) and 126 original HPV type 16 DNA copies with type-specific primers. Comparison of the amplification results for consensus L1 primers and type-specific E6-E7 primers indicated the presence of L1 deletions in 23 of 56 samples. The conclusion is that in PCR detection systems, multiple consensus primers and type-specific primers should be used in order to detect all patients harboring HPV.
机译：我们使用多种PCR引物组，试图优化从361例浸润性宫颈癌患者的冷冻活检标本中分离的DNA样品中人乳头瘤病毒（HPV）的检测。检测到的HPV分为三个不同的组，包括Telelab（挪威，斯基恩）的I / Inex组和挪威Radium医院（挪威，奥斯陆）的Ineg组和II组。共有引物组是HPV L1基因的Oli-1b-oli-2i，My09-My11，Gp5-Gp6和Gp（5 +）-Gp6 +，E1基因是CpI-CpIIG。使用这些共有引物和E6-E7的类型特异性引物，我们发现355例患者（98％）是HPV阳性。与最敏感的共有引物组相比，针对HPV 11、16、18、31、33和35型的类型特异性引物检测出更多的HPV感染患者，而三个共有引物组My，Gp / Gp +和Cp一起检测出更多HPV阳性的患者比特异性引物类型强。用连续稀释在淋巴细胞（HPV阴性细胞）中的SiHa细胞对PCR的敏感性进行测试，结果表明，使用共有引物（My，Gp +和Cp）的HPV 16型DNA拷贝的检出限为6,300，使用HPV的Cp 16型原始DNA拷贝的检出限为126。类型特异性引物。比较共有L1引物和类型特异性E6-E7引物的扩增结果表明，在56个样品中的23个中存在L1缺失。结论是在PCR检测系统中，应使用多个共有引物和特定类型的引物来检测所有携带HPV的患者。

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