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两种用于检测牛乳头瘤病毒的聚合酶链反应策略的比较。

Comparison of two PCR strategies for the detection of bovine papillomavirus.

机构信息

Department of Genetics, Federal University of Pernambuco, 50740521 Pernambuco, Brazil.

出版信息

J Virol Methods. 2013 Sep;192(1-2):55-8. doi: 10.1016/j.jviromet.2013.04.017. Epub 2013 May 10.

Abstract

Bovine papillomavirus (BPV) is a diverse group of double-stranded DNA oncogenic viruses, which have been detected in epithelial lesions and body fluids. Most studies of BPV infection rely on a single method for DNA detection; however the use of any single method or technique may underestimate the true prevalence of this virus. The purpose of this study was to compare two PCR strategies for the detection of BPV in skin lesions and fluids: these involve the use of BPV type-specific and consensus primers. Seventy-two cutaneous lesions, 57 blood samples and 59 semen samples were collected. PCR was used with the FAP consensus primers and BPV type-specific primers (for BPVs 2, 3, 4, 5, 8, 9 and 10), along with sequencing assays, to detect the BPV types. Phylogenetic analysis was carried out by means of the maximum likelihood method. It was found that both FAP and BPV type-specific primer sets could amplify BPV types of DNA in skin lesions, blood and semen samples. However, the BPV type-specific primers were more sensitive than the consensus primers and were able to detect co-infection of BPV in the samples. The consensus primers amplified five BPV types and were more suitable for detecting new putative BPV types. Thus, account should be taken of both PCR primer systems to identify co-infection, the presence of novel viruses, and avoid false-negative results.

摘要

牛乳头瘤病毒(BPV)是一组具有双链 DNA 致癌性的多样化病毒,已在上皮病变和体液中检测到。大多数 BPV 感染的研究依赖于单一的 DNA 检测方法;然而,任何单一方法或技术的使用都可能低估这种病毒的真实流行率。本研究旨在比较两种用于检测皮肤病变和体液中 BPV 的 PCR 策略:这些策略涉及使用 BPV 型特异性和通用引物。收集了 72 个皮肤病变、57 个血液样本和 59 个精液样本。使用 FAP 通用引物和 BPV 型特异性引物(用于 BPV 2、3、4、5、8、9 和 10)以及测序检测,检测 BPV 类型。通过最大似然法进行系统发育分析。结果发现,FAP 和 BPV 型特异性引物组都可以扩增皮肤病变、血液和精液样本中的 BPV 类型 DNA。然而,BPV 型特异性引物比通用引物更敏感,能够检测样本中的 BPV 混合感染。通用引物扩增了 5 种 BPV 类型,更适合检测新的潜在 BPV 类型。因此,应考虑使用两种 PCR 引物系统来识别混合感染、新型病毒的存在,并避免假阴性结果。

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