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Mouse DNA contamination in human tissue tested for XMRV

机译:XMRV检测人体组织中的小鼠DNA污染

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Background We used a PCR-based approach to study the prevalence of genetic sequences related to a gammaretrovirus, xenotropic murine leukemia virus-related virus, XMRV, in human prostate cancer. This virus has been identified in the US in prostate cancer patients and in those with chronic fatigue syndrome. However, with the exception of two patients in Germany, XMRV has not been identified in prostate cancer tissue in Europe. Most putative associations of new or old human retroviruses with diseases have turned out to be due to contamination. We have looked for XMRV sequences in DNA extracted from formalin-fixed paraffin- embedded prostate tissues. To control for contamination, PCR assays to detect either mouse mitochondrial DNA (mtDNA) or intracisternal A particle (IAP) long terminal repeat DNA were run on all samples, owing to their very high copy number in mouse cells. Results In general agreement with the US prevalence, XMRV-like sequences were found in 4.8% of prostate cancers. However, these were also positive, as were 21.5% of XMRV-negative cases, for IAP sequences, and many, but not all were positive for mtDNA sequences. Conclusions These results show that contamination with mouse DNA is widespread and detectable by the highly sensitive IAP assay, but not always with less sensitive assays, such as murine mtDNA PCR. This study highlights the ubiquitous presence of mouse DNA in laboratory specimens and offers a means of rigorous validation for future studies of murine retroviruses in human disease.
机译:背景技术我们使用基于PCR的方法研究了人类前列腺癌中与伽玛逆转录病毒,异种鼠白血病病毒相关病毒XMRV相关的基因序列的普遍性。在美国,已在前列腺癌患者和患有慢性疲劳综合症的患者中发现了这种病毒。但是,除德国的两名患者外,在欧洲的前列腺癌组织中尚未发现XMRV。事实证明,大多数新的或旧的人类逆转录病毒与疾病的关联都归因于污染。我们已经在从福尔马林固定石蜡包埋的前列腺组织中提取的DNA中寻找XMRV序列。为了控制污染,由于它们在小鼠细胞中的拷贝数非常高,因此对所有样品都进行了检测小鼠线粒体DNA(mtDNA)或脑池内A颗粒(IAP)长末端重复DNA的PCR分析。结果与美国的患病率普遍一致,在4.8%的前列腺癌中发现了XMRV样序列。但是,对于IAP序列,这些也都是阳性的,如XMRV阴性病例的21.5%一样,许多但并非全部对mtDNA序列都是阳性的。结论这些结果表明,通过高灵敏度的IAP分析可以广泛检测到小鼠DNA的污染,但并非总是通过灵敏度较低的分析(例如鼠mtDNA PCR)进行检测。这项研究突出了实验室标本中普遍存在的小鼠DNA,并为今后在人类疾病中进行鼠逆转录病毒的研究提供了严格的验证手段。

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