Mouse DNA contamination in human tissue tested for XMRV.
Mouse DNA contamination in human tissue tested for XMRV.
复制标题
DOI：
10.1186/1742-4690-7-108
复制
发表时间：
2010-12-20
期刊：
影响因子：
3.3
通讯作者：
McClure MO
中科院分区：
文献类型：
--
作者：
Robinson MJ;Erlwein OW;Kaye S;Weber J;Cingoz O;Patel A;Walker MM;Kim WJ;Uiprasertkul M;Coffin JM;McClure MO
我们使用一种基于聚合酶链式反应的方法来研究人类前列腺癌中与伽马逆转录病毒、异嗜性小鼠白血病病毒相关病毒XMRV相关的基因序列的流行率。这种病毒已经在美国前列腺癌患者和慢性疲劳综合征患者中被发现。然而，除了德国的两名患者外，欧洲尚未在前列腺癌组织中发现XMRV。大多数假定的新的或旧的人类逆转录病毒与疾病的联系被证明是由于污染。我们在从福尔马林固定的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，所有样本都进行了检测小鼠线粒体DNA(MtDNA)或脑池内A粒子(IAP)长末端重复DNA的聚合酶链式反应，因为它们在小鼠细胞中的拷贝数很高。与美国的患病率基本一致的是，4.8%的前列腺癌中发现了XMRV样序列。然而，这些也是阳性的，就像21.5%的XMRV阴性病例一样，对IAP序列也是阳性的，对mtDNA序列也有许多但不是全部阳性。这些结果表明，对小鼠DNA的污染是普遍存在的，并可通过高灵敏度的IAP检测方法检测到，但不总是使用灵敏度较低的检测方法，如小鼠线粒体DNA聚合酶链式反应。这项研究强调了实验室标本中普遍存在的小鼠DNA，并为未来人类疾病中小鼠逆转录病毒的研究提供了一种严格的验证手段。
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. 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. 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.
登录
查看更多内容
影响因子：
3.7
作者：
Erlwein O;Kaye S;McClure MO;Weber J;Wills G;Collier D;Wessely S;Cleare A
通讯作者：
Cleare A
影响因子：
3.3
作者：
Groom HC;Boucherit VC;Makinson K;Randal E;Baptista S;Hagan S;Gow JW;Mattes FM;Breuer J;Kerr JR;Stoye JP;Bishop KN
通讯作者：
Bishop KN
影响因子：
5.4
作者：
Hong, Seunghee;Klein, Eric A.;Silverman, Robert H.
通讯作者：
Silverman, Robert H.
影响因子：
56.9
作者：
Lombardi, Vincent C.;Ruscetti, Francis W.;Mikovits, Judy A.
通讯作者：
Mikovits, Judy A.
影响因子：
2.1
作者：
Arnold, Rebecca S.;Makarova, Natalia V.;Petros, John A.
通讯作者：
Petros, John A.