16S rDNA实时荧光定量PCR法与全自动培养法在血小板细菌污染检测中的应用比较  

Comparison between 16S rDNA amplification and automated culture assay in detecting bacteria contamination of platelets concentrates

作　　者：刘晓颖[1] 马敏[2] 徐忠[1] 王迅[1] 林俊杰[1] 邱颖捷[1] 钱开诚[1] 

机构地区：[1]上海市血液中心,上海200051 [2]华东师范大学

出　　处：《中国输血杂志》2008年第9期677-679,共3页Chinese Journal of Blood Transfusion

摘　　要：目的比较16S rDNA实时荧光定量PCR法与全自动培养法在血小板制品细菌污染检测中的灵敏度和特异性,评价2种方法的应用前景。方法将血小板制品污染中常见的6种细菌用浓缩血小板悬液进行稀释,并选取浓度为102、101、100的菌悬液,分别用实时荧光定量PCR法和全自动培养法进行检测。结果用16S rDNA实时荧光定量PCR法对6株细菌检测,其灵敏度和特异性均为100%,Κ=1.000。用全自动培养仪对6株细菌检测,其特异性均为100%;灵敏度分别为:金黄色葡萄球菌需氧瓶95.5%,Κ=0.886,厌氧瓶90.9%,Κ=0.787;表皮葡萄球菌及大肠杆菌2种培养瓶均为83.3%,Κ=0.667;蜡样芽孢杆菌2种培养瓶均为86.7%,Κ=0.684;铜绿假单胞杆菌需氧瓶度为100%,Κ=1.000,厌氧瓶为44.4%,Κ=0.286;痤疮丙酸杆菌需氧瓶为16.7%,Κ=0.105,厌氧瓶为91.7%,Κ=0.886。结论实时荧光定量PCR法检测血小板制品中的细菌污染,灵敏度高,特异性好,且省时、经济,能应用于临床上血液样本的大规模筛查。Objective To compare the sensitivity and specificity between 16S rDNA real-time PCR and automated culture assay in detecting bacteria contamination in platelets concentrates （ PC ）. Methods A total of 6 bacteria strains, frequently associated with the contamination of PCs, were serially diluted. Bacterial suspensions of 102, 101, and 10^0 CFU/ml were tested by real-time PCR and culture assay. Results The sensitivity and specificity of real-time PCR in detecting the 6 bacterial strains were both 100% （ K = 1. 000 ）. The specificity of automated culture system was 100%. The sensitivity of automated culture system in detecting Staphylococcus aureus in standard aerobic bottles was 95.5% （ K = 0. 886 ） , and in anaerobic bottles was 90.9% （ K = 0. 787 ）. The sensitivity in detecting Staphylococcus epidermidis and Escherichia coli were both 83.3% （ K = 0. 667 ）. The sensitivity in detecting Bacillus cereus was 86.7% （ K = 0. 684 ）. The sensitivity in detecting Pseudomonas aeruginasa were 100% （ K = 1. 000 ） in standard aerobic bottles, and 44. 4% （ K = 0. 286 } in anaerobic bottles. The sensitivity in detecting Propionibacterium acnes was 16.7% （K = 0. 105） in standard aerobic bottles, and 91.7% （K = 0. 886） in anaerobic bottles. Conclusion The real-time quantitative PCR, with high sensitivity and specificity, is a rapid and reliable method for the clinical prevention of transfusion of contaminated units.

关 键 词：血小板 细菌 实时荧光定量PCR 全自动培养 16S RDNA 

分 类 号：R457.1[医药卫生—治疗学] R446.11[医药卫生—临床医学]