真菌聚合酶链反应检测中出现的污染情况。
Contaminations occurring in fungal PCR assays.
作者信息
Loeffler J, Hebart H, Bialek R, Hagmeyer L, Schmidt D, Serey F P, Hartmann M, Eucker J, Einsele H
机构信息
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, 72076 Tuebingen, Germany.
出版信息
J Clin Microbiol. 1999 Apr;37(4):1200-2. doi: 10.1128/JCM.37.4.1200-1202.1999.
Abstract
Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified as Aspergillus fumigatus, Saccharomyces cerevisiae, and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10x PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken.
摘要
最近有报道称在临床标本中成功实现了真菌DNA的体外扩增。在欧洲五个中心的一项合作中，分析了真菌PCR中因空气传播孢子接种或残留污染导致的污染频率和风险。通过循环测序和GenBank分析确定了所有污染物的身份。在总共包含超过2800个样本的150次PCR检测中，有12次被发现受到污染（在DNA提取过程中，3.3%的阴性对照被污染，在扩增过程中，4.7%的PCR混合物被污染）。污染物被确定为烟曲霉、酿酒酵母和枝顶孢属。进一步分析表明，诸如溶菌酶粉末或10x PCR缓冲液等市售产品可能含有真菌DNA。总之，如果采取一般预防措施，真菌PCR检测中的污染风险并不高于其他基于诊断性PCR的检测。
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