实时荧光PCR快速检测创伤组织中繁殖状态金黄色葡萄球菌的方法研究

Study on a method of a rapid detection for Staphylococcus aureus from wound tissue in department of orthopedics by real-time-PCR

目的为解决临床对创口组织金黄色葡萄球菌培养鉴定时间偏长、阳性率低的不满意现象,探讨建立一种检测时间显著缩短而阳性率得到明显提高的方法。方法以金黄色葡萄球菌检测的国家标准方法为金标准,选用246份原始组织样本与其增菌后组织样本同时用实时荧光PCR法检测,与创口组织标本的金黄色葡萄球菌常规培养法作平行检测比对,经统计分析评估实时荧光PCR法和常规培养法的7项指标及标准方法的一致性与检测时间的比较。结果常规培养法与实时荧光PCR法的灵敏度分别为40.91%、100%;特异度分别为100%、100%;约登指数分别为0.41、1.00;阳性预测值分别为40.91%、100%;阴性预测值分别为88.60%、100%;与标准方法的一致性检验Kappa值分别为0.53、1.00;常规培养法的检出阳性率为7.32%,时间需3 d^4 d;实时荧光PCR法的检出阳性率为18.70%,所需时间仅为2 h^26 h。结论实时荧光PCR法的灵敏度、约登指数、检测时间和检出阳性率等均优于常规培养法,基本解决了临床对创口组织金黄色葡萄球菌培养鉴定时间偏长、阳性率低的不满意现象。

Objective In order to solve the problem of low detection rate and long culture time, we aim to investigate a rapid detection of Staphylococcus aureus from wound tissue in department of orthopedics. Methods Regard the national standard method for detection of Staphylococcus aureus as the gold standard, the 246 copy of the original tissue samples and enrichment of tissue samples were simultaneously detected by real - time fluorescence PCR for parallel detection comparison with the conven- tional culture method of Staphylococcus aureus of wound tissue samples. Through statistical analysis to evaluate the comparison of 7 indicators and standards method coincidence of real - time fluorescent the PCR method and the conventional culture method with detection time. Results Sensitivity （40.91%, 100% ）, specificity（ 100%, 100% ）, youden index （0.41,1.00）, posi- tive predictive value（40.91%, 100% ）, negative predictive value（ 88.60%, 100% ） of general bacterial culture and real -time PCR were respectively recorded. Kappa values of general bacterial culture and real - time PCR were 0.53 and 1.00. The gener- al bacterial culture need 3 d -4 d and obtained a detection rate of 7.32%. The real -time PCR need 2 h -26 h and obtain a positive detection rate of 18.70%. Conclusion real - time PCR is better than general bacterial culture in most parameters such as sensitivity, youden index, detection time and detection rate, which basically solved the problem of low detection rate and long culture time.