Parasitology-Mycology Laboratory, Bichat-Claude Bernard hospital, APHP, 75018 Paris, France - IRD MERIT UMR 261, Pharmacy Faculty, Paris University, 75006 Paris, France.
Parasitology-Mycology Laboratory, CHU de Clermont-Ferrand, 3IHP, INSERM, 63000 Clermont-Ferrand, France.
Parasite. 2022;29:5. doi: 10.1051/parasite/2022003. Epub 2022 Feb 9.
Commercial multiplex PCR assay panels were developed to overcome the limitations of microscopic examination for parasitological diagnosis on stool samples. However, given the increased supply of this diagnostic approach, these assays must be evaluated to position them in a diagnostic algorithm. Analytical performances of the multiplex PCR assay G-DiaParaTrio, Allplex GI parasite and RIDAGENE parasitic stool panel for detecting Blastocystis sp., Entamoeba histolytica, Giardia duodenalis, Cryptosporidium spp., Dientamoeba fragilis, and Cyclospora cayetanensis, were assessed through a retrospective comparative study on 184 stool samples initially sent for parasitological investigation. The composite reference method for parasitological diagnosis was microscopic observation and Entamoeba histolytica-specific adhesion detection when necessary. Multiplex PCR assays were performed on extracted DNA from each stool, following the manufacturer's recommendations. Discrepant results with the composite reference method were investigated with species-specific PCR to approach a final parasitological diagnosis. Overall sensitivity/specificity for the multiplex PCR assays was 93.2%/100% for G-DiaParaTrio, 96.5%/98.3% for Allplex GI parasite and 89.6%/98.3% for RIDAGENE, whereas the composite reference method presented an overall sensitivity/specificity of 59.6%/99.8%. These results confirmed the added diagnostic value of the multiplex PCR approach for gastrointestinal protists. Nevertheless, the PCR procedure and the analytical performance for each protist of interest, variable depending on the multiplex PCR assay, must be considered when implementing a PCR-based diagnostic approach.
商业多重 PCR 检测试剂盒的开发是为了克服粪便样本寄生虫镜检诊断的局限性。然而，鉴于这种诊断方法的供应增加，必须对这些检测方法进行评估，以便将其纳入诊断算法。通过对 184 份最初用于寄生虫学检查的粪便样本进行回顾性比较研究，评估了 G-DiaParaTrio、Allplex GI parasite 和 RIDAGENE 寄生虫粪便检测试剂盒检测蓝氏贾第鞭毛虫、溶组织内阿米巴、十二指肠贾第鞭毛虫、隐孢子虫、脆弱双核阿米巴和环孢子虫的多重 PCR 检测方法的分析性能。寄生虫学诊断的综合参考方法是必要时进行显微镜观察和溶组织内阿米巴特异性黏附检测。根据制造商的建议，对每份粪便提取的 DNA 进行多重 PCR 检测。用与综合参考方法不一致的结果进行种特异性 PCR 检测，以确定最终的寄生虫学诊断。对于 G-DiaParaTrio、Allplex GI parasite 和 RIDAGENE，多重 PCR 检测的总灵敏度/特异性分别为 93.2%/100%、96.5%/98.3%和 89.6%/98.3%，而综合参考方法的总灵敏度/特异性为 59.6%/99.8%。这些结果证实了多重 PCR 方法在胃肠道原生动物诊断中的附加诊断价值。然而，当实施基于 PCR 的诊断方法时，必须考虑到 PCR 程序和每个感兴趣的原生动物的分析性能，这取决于多重 PCR 检测方法。
