Institute of Parasitology, Biology Centre, the Czech Academy of Sciences, České Budějovice 370 05, Czech Republic - Department of Medical Biology, Faculty of Science, University of South Bohemia, České Budějovice 370 05, Czech Republic.
Institute of Parasitology, Biology Centre, the Czech Academy of Sciences, České Budějovice 370 05, Czech Republic.
Parasite. 2022;29:30. doi: 10.1051/parasite/2022029. Epub 2022 May 31.
Blastocystis is the most commonly found intestinal protist in the world. Accurate detection and differentiation of Blastocystis including its subtypes (arguably species) are essential to understand its epidemiology and role in human health. We compared (i) the sensitivity of conventional PCR (cPCR) and qPCR in a set of 288 DNA samples obtained from stool samples of gut-healthy individuals, and (ii) subtype diversity as detected by next-generation sequencing (NGS) versus Sanger sequencing. Real-time PCR resulted in more positive samples than cPCR, revealing high fecal load of Blastocystis based on the quantification curve in most samples. In subtype detection, NGS was largely in agreement with Sanger sequencing but showed higher sensitivity for mixed subtype colonization within one host. This fact together with use of the combination of qPCR and NGS and obtaining information on the fecal protist load will be beneficial for epidemiological and surveillance studies.
芽囊原虫是世界上最常见的肠道原生动物。准确检测和区分芽囊原虫（包括其亚型，可被视为种）对于了解其流行病学和在人类健康中的作用至关重要。我们比较了（i）从肠道健康个体的粪便样本中获得的 288 个 DNA 样本中常规 PCR（cPCR）和 qPCR 的灵敏度，以及（ii）下一代测序（NGS）与 Sanger 测序检测到的亚型多样性。实时 PCR 比 cPCR 产生了更多的阳性样本，根据大多数样本中的定量曲线显示出芽囊原虫的高粪便负荷。在亚型检测中，NGS 与 Sanger 测序基本一致，但在检测同一宿主内混合亚型定植方面具有更高的灵敏度。这一事实以及 qPCR 和 NGS 的联合使用和粪便原生动物负荷信息的获取将有利于流行病学和监测研究。
