Comparison of the EntericBio Multiplex PCR System with Routine Culture for Detection of Bacterial Enteric Pathogens
Comparison of the EntericBio Multiplex PCR System with Routine Culture for Detection of Bacterial Enteric Pathogens
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DOI：
10.1128/jcm.01026-09
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发表时间：
2009-11-01
影响因子：
9.4
通讯作者：
Lucey, Brigid
中科院分区：
文献类型：
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作者：
O'Leary, James;Corcoran, Daniel;Lucey, Brigid
EntericBio系统使用多重PCR方法同时检测粪便中的弯曲杆菌、肠沙门氏菌、志贺氏菌和大肠杆菌O157。它将过夜肉汤浓缩与PCR扩增和杂交检测相结合。对该体系的评价是通过将该体系所获得的结果与常规培养所获得的结果进行比较，并辅之以替代的PCR检测方法。在一项对773份样本的研究中，常规培养和EntericBio系统分别产生了94.6%和92.4%的阴性结果。有42份标本培养呈阳性，与EntericBio系统检测均为阳性。该系统还检测到17份阳性样品(弯曲杆菌属12份、志贺氏菌属1份、大肠杆菌O157属4份)，但有5份样本(弯曲杆菌属2份、志贺氏菌属1份、大肠杆菌O157属2份)的检测结果不能确定。志贺氏菌的靶标。EntericBio系统检测到的是ipah基因，是志贺氏菌存在的分子标志。经序列分析，证实该患者分离的肺炎克雷伯菌中存在ipaH基因。其敏感性、特异性、阳性预测值和阴性预测值分别为100%、99.3%、91.5%和100%。使用EntericBio系统大大缩短了周转时间，并在实验室收到样本后24至32小时内即可获得结果。此外，该系统的使用还显著减少了实验室废物量。综上所述，EntericBio系统证明使用方便，比本研究中使用的传统培养法更敏感，特异性强；而且它对所测试的病原体产生结果的速度比常规培养快得多。
The EntericBio system uses a multiplex PCR assay for the simultaneous detection of Campylobacter spp., Salmonella enterica, Shigella spp., and Escherichia coli O157 from feces. It combines overnight broth enrichment with PCR amplification and detection by hybridization. An evaluation of this system was conducted by comparing the results obtained with the system with those obtained by routine culture, supplemented with alternative PCR detection methods. In a study of 773 samples, routine culture and the EntericBio system yielded 94.6 and 92.4% negative results, respectively. Forty-two samples had positive results by culture, and all of these were positive with the EntericBio system. This system detected an additional 17 positive samples (Campylobacter spp., n = 12; Shigella spp., n = 1; E. coli O157, n = 4), but the results for 5 samples (Campylobacter spp., n = 2; Shigella spp., n = 1; E. coli O157, n = 2) could not be confirmed. The target for Shigella spp. detected by the EntericBio system is the ipaH gene, and the molecular indication of the presence of Shigella spp. was investigated by sequence analysis, which confirmed that the ipaH gene was present in a Klebsiella pneumoniae isolate from the patient. The sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 99.3%, 91.5%, and 100%, respectively. Turnaround times were significantly reduced with the EntericBio system, and a result was available between 24 and 32 h after receipt of the sample in the laboratory. In addition, the amount of laboratory waste was significantly reduced by use of this system. In summary, the EntericBio system proved convenient to use, more sensitive than the conventional culture used in this study, and highly specific; and it generated results significantly faster than routine culture for the pathogens tested.