De Rauw K, Detemmerman L, Breynaert J, Piérard D
Department of Microbiology, National Reference Centre for STEC/VTEC, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090, Brussels, Belgium.
LaCAR MDX Technologies, Liège, Belgium.
Eur J Clin Microbiol Infect Dis. 2016 Sep;35(9):1479-86. doi: 10.1007/s10096-016-2688-7. Epub 2016 Jun 3.
The purpose of this investigation was the evaluation of the performance of the BioFire FilmArray® Gastrointestinal (FA-GI) Panel, a multiplexed molecular stool screening assay, for the detection of diarrheagenic Escherichia coli (DEC), with emphasis on Shiga toxin-producing E. coli (STEC). A dilution series of 12 STEC reference strains was tested with the FA-GI Panel to assess the analytical sensitivity. A total of 389 patient samples were analyzed with the FA-GI Panel and confirmation of the detected DEC was attempted with an in-house culture-based polymerase chain reaction (PCR) method. All Shiga toxin genes, except the one encoding Stx2f, were detected in bacterial dilutions ranging from 10(4) to 10(2) colony-forming units (CFU)/ml. eae + stx2f + STEC was misclassified as enteropathogenic E. coli (EPEC). Different sensitivities for various gene targets present in one isolate led to differing identifications depending on the concentration. Using the in-house method as a reference, the FA-GI Panel had a sensitivity of 90.6 % [confidence interval (CI) 75.0 %-98.0 %] and a specificity of 97.2 % (CI 94.9 %-98.6 %) for STEC detection in feces. At least one DEC was reported in 35.5 % (171/389) of the patient specimens, with EPEC being the most prevalent (n = 71). Only 59.7 % of the detected DEC could be confirmed, presumably because the comparator method was not applied directly on feces. The FA-GI Panel could not detect the stx2f subtype, misclassified certain pathogens, and the high detection rate of EPEC needs further investigation. Nevertheless, we believe that this sensitive and convenient system will prove to be an invaluable tool for the rapid diagnosis of most DEC infections, but culturing of the detected microorganisms should always be attempted.
本研究的目的是评估BioFire FilmArray®胃肠道(FA-GI)检测板,一种多重分子粪便筛查检测方法,用于检测致泻性大肠杆菌(DEC)的性能,重点是产志贺毒素大肠杆菌(STEC)。用FA-GI检测板检测了12株STEC参考菌株的稀释系列,以评估分析灵敏度。用FA-GI检测板分析了389份患者样本,并尝试用基于培养的内部聚合酶链反应(PCR)方法确认检测到的DEC。除了编码Stx2f的基因外,所有志贺毒素基因在10⁴至10²菌落形成单位(CFU)/ml的细菌稀释液中均能被检测到。eae⁺stx2f⁺ STEC被误分类为肠致病性大肠杆菌(EPEC)。同一分离株中存在的各种基因靶点的不同敏感性导致根据浓度不同而有不同的鉴定结果。以内部方法为参考,FA-GI检测板对粪便中STEC检测的灵敏度为90.6%[置信区间(CI)75.0%-98.0%],特异性为97.2%(CI 94.9%-98.6%)。在35.5%(171/389)的患者标本中报告了至少一种DEC,其中EPEC最为常见(n = 71)。仅59.7%检测到的DEC能够得到确认,可能是因为比较方法未直接应用于粪便。FA-GI检测板无法检测到stx2f亚型,对某些病原体进行了错误分类以及EPEC的高检出率需要进一步研究。然而,我们认为这个灵敏且便捷的系统将被证明是快速诊断大多数DEC感染的宝贵工具,但始终应尝试对检测到的微生物进行培养。