多重聚合酶链技术诊断重症急性胰腺炎继发感染的临床价值被引量：4

Clinical value of multiplex PCR in the diagnosis of secondary infection of severe acute pancreatitis

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摘要目的探讨多重聚合酶链反应技术（m—PCR）对重症急性胰腺炎（SAP）继发感染的诊断价值，为临床SAP抗感染治疗提供依据。方法选取2011年1月至2014年12月期间收治的35例SAP患者，在患者发病7～14d内抽外周血使用m—PCR法检测患者有无继发性细菌感染，同时抽取外周血和（或）取CT引导下腹腔穿刺液进行培养，以血培养或穿刺液培养阳性为诊断继发性感染的标准。结果采用m—PCR可同时检测9种肠道常驻致病菌，检测下限为10～1000个拷贝。培养法及m—PCR法检出金黄色葡萄球菌分别为6例和5例，表皮葡萄球菌11例和9例，粪肠球菌2例和3例，屎肠球菌6例和7例，大肠埃希菌19例和17例，肺炎克雷伯菌例2例和3例，铜绿假单胞菌6例和4例，鲍曼不动杆菌2例和2例，嗜麦芽窄食单胞菌4例和2例。35例SAP患者在发病后7—14d内27例血培养或穿刺液培养阳性，8例阴性。30例m—PCR检测阳性，5例阴性。3例培养阴性者m—PCR为阳性，其余32例两种检测方法结果一致。以培养结果为准，m—PCR的灵敏性为100％，特异性为62．5％，准确性为91．43％，阳性预测值90％，阴性预测值100％。血液或穿刺液从培养到出报告时间为（26±15）h，到鉴定结果的报告时间为（102±32）h；m—PCR方法从检测到出报告时间为（12±8）h。m—PCR检出病原菌的报告时间明显短于传统培养与鉴定结果报告时间，组间差异有统计学意义（P〈0．05）。结论m—PCR可用于监测SAP患者继发细菌感染，其敏感性高，操作时间短，值得临床推广应用。 Objective To investigate the diagnostic value of multiplex polymerase chain reaction （m-PCR） for diagnosing second infection of severe acute pancreatitis （SAP）, and to provide evidence for anti-infection treatment of SAP. Methods From January 2011 to December 2014, thirty five patients of SAP were enrolled. Seven to fourteen days after SAP onset, patients＇ blood samples were taken and the presence of infection was determined by m-PCR. In the meantime, peripheral blood or the paracentesis fluid was cultured, and the result of culture was regard as golden standard to diagnose infection. Results The m-PCR could simultaneously detect 9 kinds of intestinal resident pathogenic bacteria, and the lower limit of detection was 10 - 1 000 copies. The detection rates were as follows （cultivation vs. m-PCR）： staphylococcus aureus 6 vs 5 cases, staphylococcus epidermidis 11 vs 9 cases, enterocoecus faecalis 2 vs 3 cases, enterocoecus faecium 6 vs 7 cases, escherichia coli 19 vs 17 cases, klebsiella pneumoniae 2 vs 3 cases, pseudomonas aeruginosa 6 vs 4 cases, acinetobacter baumannii 2 vs 2 cases, malt narrow food aeromonas 4 vs 2 cases. The 7th - 14th days after SAP onset, the blood or paracentesis fluid culture was positive in 27 patients, and negative in 8 cases. And the m-PCR results were positive in 30 patients, and negative in 8 cases. The m-PCR results were positive in 30 patients, negative in 5 patients. The m-PCR results were positive in 3 patients who had negative culture results. In the remaining 32 cases, the results were consistent between the two detection methods. When the culture result was regarded as golden standard, the sensitivity, specificity and accuracy of m-PCR were 100%, 62.5% and 91.43% , respectively. The positive predictive value and the negative predictive value were 90% and 100%, respectively. It took （26 ± 15） hours on average to obtain the result of culture method, and it took （102 ±32） hours on average to obtain the confirmative results. It took （12 ± 8） hours on average to obtain the result of the m-PCR method. The time course of m-PCR was significantly shorter than that of the traditional culture method, and the difference was statistically significant （ P 〈 0.05 ）. Conclusions The m-PCR method can be used to monitor the bacterial infection in patients with SAP. The m-PCR method is a highly sensitive and rapid detection approach, which is worth of clinical application.