16S rRNA通用引物在血小板制品细菌污染检测中的应用  

Application of 16S ribosomal RNA general primer in the bacterial detection of platelet concentrates

作　　者：孟庆丽[1] 黄杰[2] 于卫建[1] 叶萍[1] 席惠君[1] 

机构地区：[1]大连市红十字血液中心,辽宁大连116001 [2]大连医科大学检验医学院

出　　处：《中国输血杂志》2008年第5期349-351,共3页Chinese Journal of Blood Transfusion

摘　　要：目的评价16S rRNA通用引物在快速检出血小板制品中污染细菌的实用性,寻找新的能够快速、准确地检出血小板中污染细菌的方法。方法用分子生物学方法提取样本中细菌DNA,用设计合成的16S rRNA和16S—23S rRNA基因区间引物进行扩增,产物经HaeⅢ酶切,酶切片段与血小板污染常见菌的酶切图谱对比,确定有无污染及污染菌种类。结果16S rRNA通用引物法可以在4h内完成全部检测,其中在保存24h的血小板标本中未检出污染菌,保存48h的血小板标本中有2份分别检出了金黄色葡萄球菌和枯草芽孢杆菌,与全自动细菌培养仪的结果一致。结论16S rRNA通用引物是一种快速、准确检出血小板制品中污染细菌的方法,结果可靠,具有较高的临床应用价值。Objective To evaluate the effectiveness of 16S ribosomal RNA（16S rRNA） general primer in bacterial detection and to find a method that could detect the bacteria rapidly and accurately in platelet concentrates. Methods Sample Genomic DNA was extracted by molecular biology method. 16S rRNA general primer and 16S-23S ribosomal RNA interval primer were used for PCR, and the PCR products were treated with enzyme Hae Ⅲ. We compared the enzyme treated fragments with frequent bacteria sheet to determine whether the sample was contaminated or not. Once the sample was contaminated, the contaminated strain had to been confirmed. Results All detection were finished within 4 hours with 16S rRNA general primer method. No bacterium-was detected in platelet concentrates during the 24-hour preservation. While in platelet concentrates preserved for 48-hour, S. epiderimidis and B. subtilis were detected, and the results were confirmed through auto bacteria culture. Conclusion 16S rRNA general primer method is rapid and accurate in the bacterial detection of platelet concentrates, suggesting the reliability and clinical value.

关 键 词：16S RRNA 血小板 细菌 

分 类 号：R446[医药卫生—诊断学] R372[医药卫生—临床医学]