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首页> 外文期刊>Revista Argentina de Microbiologia >Detection of toxigenic Clostridioides [Clostridium] difficile: Usefulness of two commercially available enzyme immunoassays and a PCR assay on stool samples and stool isolates
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Detection of toxigenic Clostridioides [Clostridium] difficile: Usefulness of two commercially available enzyme immunoassays and a PCR assay on stool samples and stool isolates

机译：产毒性艰难梭菌[Clostridium]艰难梭菌的检测：两种市售酶免疫测定以及粪便样品和粪便分离株的PCR测定的实用性

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The best laboratory diagnostic approach to detect Clostridioides [Clostridium] difficile infection (CDI) is a subject of ongoing debate. With the aim of evaluating four laboratory diagnostic methods, 250 unformed stools from patients with suspected CDI submitted to nine medical center laboratories from November 2010 to December 2011, were studied using: (1) an immunochromatographic rapid assay test that combines the qualitative determination of glutamate dehydrogenase (GDH) plus toxins A and B (QAB), the CDIFF QUIK CHEK COMPLETE assay; (2) an enzyme immunoassay for qualitative determination of toxins A and B, the RIDASCREEN? C. difficile Toxin A/B assay (RAB); (3) a PCR for the toxin B gene assay (PCR); and (4) the toxigenic culture (TC). C. difficile isolates from direct toxin negative stools by QAB, RAB and PCR were evaluated for toxigenicity by the same direct tests, in order to assess the contribution of the TC (QAB-TC, RAB-TC, PCR-TC). A combination of the cell culture cytotoxicity neutralization assay (CCCNA) in stools, and the same assay on isolates from direct negative samples (CCCNA-TC) was considered the reference method (CCCNA/CCCNA-TC). Of the 250 stools tested, 107 (42.8%) were positive by CCCNA/CCCNA-TC. The GDH and PCR/PCR-TC assays were the most sensitive, 91.59% and 87.62%, respectively. The QAB, RAB, QAB/QAB-TC and RAB/RAB-TC had the highest specificities, ca. 95%. A negative GDH result would rule out CDI, however, its low positive likelihood ratio (PLR) of 3.97 indicates that a positive result should always be complemented with the detection of toxins. If the RAB, QAB, and PCR assays do not detect toxins from direct feces, the toxigenic culture should be performed. In view of our results, the most accurate and reliable methods to be applied in a clinical microbiology laboratory were the QAB/QAB-TC, and RAB/RAB-TC, with PLRs >10 and negative likelihood ratios <0.30.
机译：最好的实验室诊断方法来检测艰难梭菌（Clostridium）艰难梭菌感染（CDI）是一个不断争论的话题。为了评估四种实验室诊断方法，研究了从2010年11月至2011年12月向9个医疗中心实验室提交的250份疑似CDI患者未形成的粪便，方法是：（1）结合了谷氨酸的定性检测的免疫色谱快速测定法脱氢酶（GDH）以及毒素A和B（QAB），CDIFF QUICK CHEK COMPLETE分析; （2）用于定性测定毒素A和B的酶免疫测定法，RIDASCREEN？艰难梭菌毒素A / B测定（RAB）; （3）用于毒素B基因测定的PCR（PCR）; （4）产毒文化（TC）。通过QAB，RAB和PCR从直接毒素阴性粪便中分离的艰难梭菌通过同样的直接测试评估其毒性，以评估TC的贡献（QAB-TC，RAB-TC，PCR-TC）。粪便中细胞培养物细胞毒性中和测定（CCCNA）和直接阴性样品分离物（CCCNA-TC）的相同测定相结合被认为是参考方法（CCCNA / CCCNA-TC）。在250份测试的粪便中，有CCCNA / CCCNA-TC阳性107份（42.8％）。 GDH和PCR / PCR-TC检测最敏感，分别为91.59％和87.62％。 QAB，RAB，QAB / QAB-TC和RAB / RAB-TC具有最高的特异性。 95％。 GDH阴性会排除CDI，但是其低的3.97阳性似然比（PLR）表示阳性结果应始终与毒素检测相辅相成。如果RAB，QAB和PCR分析未检测到直接粪便中的毒素，则应进行产毒培养。根据我们的结果，在临床微生物学实验室中应用的最准确，最可靠的方法是QAB / QAB-TC和RAB / RAB-TC，PLR> 10，负似然比<0.30。

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