Beutin L, Steinrück H, Krause G, Steege K, Haby S, Hultsch G, Appel B
National Reference Laboratory for Escherichia coli, Federal Institute for Risk Assessment BfR, Berlin, Germany.
J Appl Microbiol. 2007 Mar;102(3):630-9. doi: 10.1111/j.1365-2672.2006.03139.x.
To evaluate the suitability of the commercially distributed Ridascreen Verotoxin enzyme immunoassay (EIA) for detection of known genetic types of the Vero (Shiga) toxins 1 (Stx1) and 2 (Stx2) families and to determine its relative sensitivity and specificity.
The Ridascreen-EIA was compared with the Vero cell assay, a P(1)-glycoprotein receptor EIA and with stx gene-specific PCs for detection of Stx with 43 Shiga toxin-producing strains of Escherichia coli (STEC) reference strains and with 241 test strains. The Ridascreen-EIA detects strains producing Stx1 and variants Stx1c and Stx1d, as well as Stx2 and variants Stx2d1, Stx2d2, Stx2e, Stx2d, Stx2-O118 (Stx2d-ount), Stx2-NV206, Stx2f and Stx2g. The assay showed a relative sensitivity of 95.7% and a relative specificity of 98.7%. Some of the Stx2-O118-, Stx2e- and Stx2g-producing STEC were not detected with the Ridascreen-EIA probably because of low amount of toxin produced by these strains.
The Ridascreen-EIA is able to detect all known types of Stx and is applicable for routine screening of bacterial isolates owing to its high specificity. It is less applicable for testing samples where low amounts of Stx are expected, such as mixed cultures and certain Stx2 variants.
This study presents a first comprehensive evaluation of the Ridascreen-EIA, a rapid standardized STEC screening test for routine diagnostic laboratories. Data are presented on the type of the spectrum of Stx that are detected with this immunoassay and its advantages and limits for practical use.
评估市售的Ridascreen Verotoxin酶免疫测定法(EIA)用于检测已知基因类型的Vero(志贺)毒素1(Stx1)和2(Stx2)家族的适用性,并确定其相对灵敏度和特异性。
将Ridascreen-EIA与Vero细胞测定法、P(1)-糖蛋白受体EIA以及stx基因特异性聚合酶链反应(PCR)进行比较,以检测43株产志贺毒素大肠杆菌(STEC)参考菌株和241株测试菌株中的Stx。Ridascreen-EIA可检测产生Stx1及其变体Stx1c和Stx1d,以及Stx2及其变体Stx2d1、Stx2d2、Stx2e、Stx2d、Stx2-O118(Stx2d-ount)、Stx2-NV206、Stx2f和Stx2g的菌株。该测定法的相对灵敏度为95.7%,相对特异性为98.7%。一些产生Stx2-O118、Stx2e和Stx2g的STEC未被Ridascreen-EIA检测到,可能是因为这些菌株产生的毒素量较低。
Ridascreen-EIA能够检测所有已知类型的Stx,因其高特异性适用于细菌分离株的常规筛查。它不太适用于检测预期Stx含量较低的样品,如混合培养物和某些Stx2变体。
本研究首次对Ridascreen-EIA进行了全面评估,这是一种用于常规诊断实验室的快速标准化STEC筛查试验。给出了用这种免疫测定法检测到的Stx谱类型及其实际应用的优点和局限性的数据。