School of Life Sciences, University of Technology Sydney, Broadway, NSW, Australia.
Division of Microbiology, Sydpath, St Vincent's Hospital, Darlinghurst, NSW, Australia
J Clin Microbiol. 2019 Apr 26;57(5). doi: 10.1128/JCM.01466-18. Print 2019 May.
is a gastrointestinal trichomonad parasite whose pathogenicity is yet to be determined. The difficulty involved in microscopically diagnosing in feces led to the development of real-time PCR methodologies for the detection of in stool samples. Prevalence studies in Europe show much higher levels of infection where a laboratory-developed real-time assay is the predominant assay for the detection of than in regions that use the EasyScreen assay for detection of gastrointestinal pathogens. The aim of this study was to compare a commercially available assay (Genetic Signatures EasyScreen assay) to a widely used laboratory-developed real-time PCR method. Two hundred fifty fecal samples were screened using the laboratory-developed real-time assay on four real-time PCR platforms producing a number of discrepant results. Limit-of-detection studies were undertaken to attempt to resolve sensitivity for each platform tested. The presence or absence of DNA in discrepant samples was shown using PCR amplicon next-generation sequencing. Eukaryotic 18S diversity profiling was conducted on discrepant samples to identify the presence or absence of additional protozoan species in samples that may be responsible for cross-reactivity seen in these samples. The results revealed the potential for multiple false-positive results when using the laboratory-developed real-time assay across multiple real-time platforms using manufacturer default settings. This report provides recommendations to resolve these issues where possible and suggestions for future prevalence studies, and it emphasizes the EasyScreen assay as the molecular method of choice as well as the need for standardization of detection assays across all nations screening for .
是一种胃肠道毛滴虫寄生虫，其致病性尚未确定。由于在粪便中显微镜诊断 存在困难，因此开发了实时 PCR 方法来检测粪便样本中的 。欧洲的流行率研究表明，在使用实验室开发的实时检测方法检测 的地区，感染水平要高得多，而在使用 EasyScreen 检测胃肠道病原体的地区则更高。本研究旨在比较一种商业上可用的 检测方法（Genetic Signatures EasyScreen 检测法）和一种广泛使用的实验室开发的实时 PCR 方法。使用实验室开发的实时 PCR 方法在四个实时 PCR 平台上筛选了 250 个粪便样本，产生了许多不一致的结果。进行了最低检测限研究，试图解决每个测试平台的灵敏度问题。使用 PCR 扩增子下一代测序显示了差异样本中 是否存在 DNA。对差异样本进行真核生物 18S 多样性分析，以确定在这些样本中可能导致交叉反应的其他原生动物物种的存在或不存在。结果表明，在使用实验室开发的实时检测法在多个实时平台上使用制造商默认设置时，可能会出现多个假阳性结果。本报告提供了尽可能解决这些问题的建议，并对未来的流行率研究提出了建议，同时强调了 EasyScreen 检测法作为首选的分子检测方法，以及在所有筛查 的国家中检测方法标准化的必要性。
