核酸扩增法检测细菌的方法研究及其在浓缩血小板制品细菌检测中的初步应用  

The study on nucleic acid testing for bacterial detection and its preliminary application on concentrated platelet

作　　者：刘宇宁 王迅[2] 费丹娜 郝旭赞 符沙鸥 贾尧[2] 

机构地区：[1]上海市奉贤区血站,上海201400 [2]上海市血液中心 [3]深圳爱康生物科技有限公司

出　　处：《中国输血杂志》2017年第7期702-704,共3页Chinese Journal of Blood Transfusion

基　　金：上海市奉贤区科学技术发展基金(奉科20141110);第四轮上海市公共卫生三年行动计划重点学科建设项目-<输血医学>(15GWZK0501)

摘　　要：目的建立符合自动化检测要求的核酸扩增法检测细菌的方法,并初步应用于手工浓缩血小板制品的细菌检测。方法对细菌DNA提取方法进行自动化改造,使用不同菌株评估改良后的方法的灵敏度和特异性,并将该方法应用于100例手工浓缩血小板的检测。结果对细菌DNA提取方法进行改造,满足了自动化检测的要求,该方法对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌和铜绿假单胞菌的灵敏度分别可达10、150、65、15 CFU/m L。使用灭菌水检测50次,均未发现非特异性检测产物。该方法和细菌培养方法同时能从100例手工浓缩血小板中检出1例细菌阳性标本。结论细菌基因组16S DNA检测方法能快速灵敏地检出血小板制品中常见的污染细菌,应用于手工浓缩血小板检测能获得与细菌培养相同的效果。Objective To establish a nucleic acid testing assay for bacterial detection with meeting automatic require- ment, and use it preliminarily to detect bacterial contamination in concentrated platelet. Methods Optimizing the bacterial DNA extraction methods, evaluating the sensitivity and specificity of the optimized assay with different bacterial strains, and applying the assay on bacterial detection for 100 units of concentrated platelet. Results The optimized extraction methods met the criteria of automatic testing. The sensitivity of the assay is 10,150,65,15 CFU/mL for Escheriehia coli, Stphylococ- cus aureus, Bacillus subtilis, and Pseudomonas aeruginas respectively. There was no non-specific amplification detected. And the assay could detect 1 contaminated bacterial from 100 units of concentrated platelet as same as BactALT bacterial cul- ture. Conclusions Nucleic acid testing for bacterial genome 16S DNA could detect common contaminated bacterial sensi- tively and fast from platelet product. This assay could have the same effect when compared to bacterial culture methods when it was applied in concentrated platelet products.

关 键 词：细菌污染 浓缩血小板 核酸检测 自动化检测 

分 类 号：R446.1[医药卫生—诊断学] R457.14[医药卫生—临床医学]