DNA extraction columns contaminated with murine sequences.

DNA extraction columns contaminated with murine sequences.
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DOI:
10.1371/journal.pone.0023484
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发表时间:
2011
期刊:
影响因子:
3.7
通讯作者:
McClure MO
McClure MO
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Erlwein O;Robinson MJ;Dustan S;Weber J;Kaye S;McClure MO

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新型伽马逆转录病毒、异种鼠白血病病毒相关病毒 (XMRV) 的序列已在人类前列腺癌组织中得到描述,尽管 DNA 含量较低。此外,据报道,慢性疲劳综合征 (CFS) 患者中存在 XMRV 序列和多向 (p) 鼠白血病病毒 (MLV)。在评估 XMRV 在前列腺癌组织样本中的流行情况时,我们发现,当使用设计用于检测小鼠基因组 DNA 污染的引物进行 PCR 时,从原始 DNA 纯化柱中洗脱出来的 XMRV 偶尔会产生扩增产物。使用引物检测 XMRV 的进一步 PCR 分析揭示了源自小鼠和人类 DNA 以及来源​​不明的 DNA 的 XMRV 和 pMLV 的序列。因此,当使用高灵敏度扩增技术检测微量 DNA 靶标时,DNA 纯化柱可能会出现问题。
Sequences of the novel gammaretrovirus, xenotropic murine leukemia virus-related virus (XMRV) have been described in human prostate cancer tissue, although the amounts of DNA are low. Furthermore, XMRV sequences and polytropic (p) murine leukemia viruses (MLVs) have been reported in patients with chronic fatigue syndrome (CFS). In assessing the prevalence of XMRV in prostate cancer tissue samples we discovered that eluates from naïve DNA purification columns, when subjected to PCR with primers designed to detect genomic mouse DNA contamination, occasionally gave rise to amplification products. Further PCR analysis, using primers to detect XMRV, revealed sequences derived from XMRV and pMLVs from mouse and human DNA and DNA of unspecified origin. Thus, DNA purification columns can present problems when used to detect minute amounts of DNA targets by highly sensitive amplification techniques.
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