通过5'核酸酶PCR检测法快速鉴定血液中的小肠结肠炎耶尔森菌。
Rapid identification of Yersinia enterocolitica in blood by the 5' nuclease PCR assay.
作者信息
Sen K
机构信息
Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland 20852, USA.
出版信息
J Clin Microbiol. 2000 May;38(5):1953-8. doi: 10.1128/JCM.38.5.1953-1958.2000.
Abstract
Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5' nuclease TaqMan PCR assay was developed to detect Y. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolitica cells, and total chromosomal DNA was extracted. When the TaqMan PCR assay was performed, as few as six bacteria spiked in 200 microliter of blood could be detected. The assay was specific and did not detect other Yersinia species. The TaqMan assay is easy to perform, takes 2 h, and has the potential for use in the rapid detection of Y. enterocolitica contamination in stored blood units.
摘要
小肠结肠炎耶尔森菌占因输注受污染红细胞导致的临床败血症发作病例的50%。开发了一种5'核酸酶TaqMan PCR检测法来检测血液中的小肠结肠炎耶尔森菌。基于小肠结肠炎耶尔森菌16S rRNA基因的核苷酸序列设计了引物和探针。向全血样本中加入不同数量的小肠结肠炎耶尔森菌细胞，然后提取总染色体DNA。进行TaqMan PCR检测时，在200微升血液中加入低至6个细菌都能被检测到。该检测法具有特异性，不会检测到其他耶尔森菌属物种。TaqMan检测法操作简便，耗时2小时，有潜力用于快速检测储存血液单位中的小肠结肠炎耶尔森菌污染情况。
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