Identification of the most common pathogenic bacteria in patients with suspected sepsis by multiplex PCR
Identification of the most common pathogenic bacteria in patients with suspected sepsis by multiplex PCR
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DOI：
10.3855/jidc.3856
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发表时间：
2014-04-01
影响因子：
1.9
通讯作者：
Esfahani, Bahram Nasr
中科院分区：
文献类型：
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作者：
Arabestani, Mohammad Reza;Fazzeli, Hossein;Esfahani, Bahram Nasr
简介：金黄色葡萄球菌、凝固酶阴性葡萄球菌、肠球菌属，已发现肠杆菌科、铜绿假单胞菌和鲍曼不动杆菌是败血症患者中最常见的菌血症致病菌。血流感染（BSI）的早期检测在临床环境中至关重要。与常规微生物学方法并行开发了一种用于鉴定临床样本中这些病原体的多重PCR方法。方法：为每种生物体选择的靶基因对于设计引物非常特异。使用Mega 4、Allel ID 6、Oligo 6和Oligo分析仪软件进行引物设计。结果：与BC相比，通用PCR和多重PCR的敏感性和特异性分别为83.87%和91.58%;与BC相比，通用PCR和多重PCR的敏感性和特异性分别为74.19%和91.58%;两种PCR的阳性预测值（PPV）和阴性预测值（NPV）分别为76.47%和94.57%、74.19%和91.58%。PCR未能鉴定细菌，而常规通用和多重PCR仅在3.96%和6.34%的病例中发现细菌（大多数细菌未包括在PCR盒中）。在6.34%的情况下，多重PCR提供了细菌鉴定，但BC显示没有细菌的samples.Conclusions：多重PCR方法有利于在几个小时内检测菌血症的血液样本。通过多重PCR快速检测细菌似乎是一种有价值的工具，可以在危重患者中早期采用病原体的抗菌治疗。
Introduction: Staphylococcus aureus, coagulase-negative staphylococci, Enterococcus spp., Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumanii have been found to be the most prevalent bacteremia-causing bacteria in patients with septicemia. Early detection of bloodstream infection (BSI) is crucial in the clinical setting. A multiplex PCR method for identification of these agents in clinical samples has been developed in parallel by conventional microbiological methods.Methodology: The target genes selected for each of the organisms were very specific for designing primers. Design of primers was done using Mega4, Allel ID6, Oligo6, and Oligo analyzer software. The test comprises a universal PCR from the 16S rDNA gene and multiplex PCR from the rpoB, gyrA, sss, and chromosome X (as an internal control).Results: The sensitivity and specificity for universal PCR and multiplex PCR in comparison with BC were 83.87% and 91.58%, and 74.19% and 91.58%, respectively. The positive predictive value (PPV) and the negative predictive value (NPV) for these two PCRs were 76.47% and 94.57%, and 74.19% and 91.58%, respectively. PCR failed to identify bacteria which were found conventionally in only 3.96% and 6.34% of the cases by universal and multiplex PCR (mostly bacteria not included in the PCR cassette). In 6.34% of the cases, multiplex PCR afforded identification of bacteria, but BC showed no bacteria in the sample.Conclusions: The multiplex PCR approach facilitates the detection of bacteremia in blood samples within a few hours. Rapid detection of bacteria by multiplex PCR appears to be a valuable tool, allowing earlier pathogen-adopted antimicrobial therapy in critically ill patients.