Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Madrid, Spain.
Department of Microbiology, Paediatrics, Radiology, and Public Health, Faculty of Medicine, University of Zaragoza, Saragossa, Spain.
Microbiol Spectr. 2022 Jun 29;10(3):e0053122. doi: 10.1128/spectrum.00531-22. Epub 2022 May 3.
spp., Giardia duodenalis, and Entamoeba histolytica are the most common diarrhea-causing protozoan species globally. Misdiagnosis is a concern for asymptomatic and chronic infections. Multiplexing, i.e., the detection of more than one parasite in a single test by real-time PCR, allows high diagnostic performance with favorable cost-effectiveness. We conducted a clinical evaluation of the VIASURE , Giardia, & E. histolytica real-time PCR assay (CerTest Biotec, San Mateo de Gállego, Spain) against a large panel ( = 358) of well-characterized DNA samples positive for spp. ( = 96), G. duodenalis ( = 115), E. histolytica ( = 25), and other parasitic species of the phyla Amoebozoa ( = 11), Apicomplexa ( = 14), Euglenozoa ( = 8), Heterokonta ( = 42), Metamonada ( = 37), Microsporidia ( = 4), and Nematoda ( = 6). DNA samples were obtained from clinical stool specimens or cultured isolates in a national reference center. Estimated sensitivity and specificity were 0.96 and 0.99 for spp., 0.94 and 1 for G. duodenalis, and 0.96 and 1 for E. histolytica, respectively. Positive and negative predictive values were calculated as 1 and 0.98 for spp., 0.99 and 0.98 for G. duodenalis, and 1 and 0.99 for E. histolytica, respectively. The assay identified six species (Cryptosporidium hominis, Cryptosporidium parvum, Cryptosporidium canis, Cryptosporidium felis, Cryptosporidium scrofarum, and Cryptosporidium ryanae) and four G. duodenalis assemblages (A, B, C, and F). The VIASURE assay provides rapid and accurate simultaneous detection and identification of the most commonly occurring species and genetic variants of diarrhea-causing parasitic protozoa in humans. Thorough independent assessment of the diagnostic performance of novel diagnostic assays is essential to ascertain their true usefulness and applicability in routine clinical practice. This is particularly true for commercially available kits based on multiplex real-time PCR aimed to detect and differentiate multiple pathogens in a single biological sample. In this study, we conducted a clinical evaluation of the VIASURE , Giardia, & E. histolytica real-time PCR assay (CerTest Biotec) for the detection and identification of the diarrhea-causing enteric protozoan parasites spp., G. duodenalis, and E. histolytica. A large panel of well-characterized DNA samples from clinical stool specimens or cultured isolates from a reference center was used for this purpose. The VIASURE assay demonstrated good performance for the routine testing of these pathogens in clinical microbiological laboratories.
spp.、十二指肠贾第鞭毛虫和溶组织内阿米巴是全球最常见的引起腹泻的原生动物物种。无症状和慢性感染存在误诊的问题。多重检测，即在实时 PCR 中一次检测一种以上寄生虫，可以提高诊断性能，具有良好的成本效益。我们对 VIASURE 、贾第鞭毛虫和溶组织内阿米巴实时 PCR 检测（西班牙圣马蒂尔德加尔莱戈的 CerTest Biotec）进行了临床评估，该检测针对的是一个由 358 个经过充分特征描述的 DNA 样本组成的大样本，这些样本对 spp.（96 个）、十二指肠贾第鞭毛虫（115 个）、溶组织内阿米巴（25 个）和其他原生动物门的寄生虫种属呈阳性，包括顶复门（11 个）、孢子虫门（14 个）、动鞭毛门（8 个）、异鞭毛门（42 个）、肉足鞭毛门（37 个）、微孢子虫门（4 个）和线虫门（6 个）。DNA 样本来自临床粪便标本或国家参考中心的培养分离物。估计的敏感性和特异性分别为 spp.（0.96 和 0.99）、十二指肠贾第鞭毛虫（0.94 和 1）和溶组织内阿米巴（0.96 和 1）。阳性和阴性预测值分别为 spp.（1 和 0.98）、十二指肠贾第鞭毛虫（0.99 和 0.98）和溶组织内阿米巴（1 和 0.99）。该检测可以识别六种 种属（人隐孢子虫、微小隐孢子虫、犬隐孢子虫、猫隐孢子虫、猪隐孢子虫和瑞安隐孢子虫）和四种十二指肠贾第鞭毛虫聚集群（A、B、C 和 F）。VIASURE 检测提供了快速、准确的同时检测和鉴定人类腹泻性寄生虫原生动物中最常见的种属和遗传变异。对新型诊断检测的诊断性能进行全面的独立评估对于确定其在常规临床实践中的真正有用性和适用性至关重要。对于基于多重实时 PCR 的商业试剂盒尤其如此，这些试剂盒旨在检测和区分单个生物样本中的多种病原体。在这项研究中，我们对 VIASURE 、贾第鞭毛虫和溶组织内阿米巴实时 PCR 检测（CerTest Biotec）进行了临床评估，用于检测和鉴定引起腹泻的肠原生动物寄生虫 spp.、十二指肠贾第鞭毛虫和溶组织内阿米巴。为此，使用了来自参考中心的临床粪便标本或培养分离物的大量经过充分特征描述的 DNA 样本。VIASURE 检测在临床微生物学实验室中对这些病原体的常规检测具有良好的性能。
