Binder Ramona, Hahn Andreas, Eberhardt Kirsten Alexandra, Hagen Ralf Matthias, Rohde Holger, Loderstädt Ulrike, Feldt Torsten, Sarfo Fred Stephen, Di Cristanziano Veronica, Kahlfuss Sascha, Frickmann Hagen, Zautner Andreas Erich
Laboratory Department, Bundeswehr Hospital Hamburg, 20359 Hamburg, Germany.
Institute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, Germany.
Microorganisms. 2023 May 17;11(5):1313. doi: 10.3390/microorganisms11051313.
Potential etiological relevance for gastroenteric disorders including diarrhea has been assigned to . However, standard routine diagnostic algorithms for stool samples of patients with diarrhea are rarely adapted to the detection of this pathogen and so, is likely to go undetected unless it is specifically addressed, e.g., by applying pathogen-specific molecular diagnostic approaches. In the study presented here, we compared three real-time PCR assays targeting the genes , (both hybridization probe assays) and (fluorescence resonance energy transfer assay) of in a test comparison without a reference standard using a stool sample collection with a high pretest probability from the Ghanaian endemicity setting. Latent class analysis was applied with the PCR results obtained with a collection of 1495 stool samples showing no signs of PCR inhibition to assess the real-time PCR assays' diagnostic accuracy. Calculated sensitivity and specificity were 93.0% and 96.9% for the -PCR, 100% and 98.2% for the -PCR, as well as 12.7% and 99.8% for the -PCR, respectively. The calculated prevalence within the assessed Ghanaian population was 14.7%. As indicated by test results obtained with high-titer spiked samples, cross-reactions of the -assay and -assay with phylogenetically related species such as can occur but are less likely with phylogenetically more distant species like, e.g., . In conclusion, the -assay showed the most promising performance characteristics as the only assay with sensitivity >95%, albeit associated with a broad 95%-confidence interval. In addition, this assay showed still-acceptable specificity of >98% in spite of the known cross-reactivity with phylogenetically closely related species such as . If higher certainty is desired, the -assay with specificity close to 100% can be applied for confirmation testing with samples showing positive -PCR results. However, in case of a negative result in the -assay, this cannot reliably exclude the detection of in the -assay due to the -assay's very low sensitivity.
腹泻等胃肠疾病的潜在病因相关性已归因于……。然而，腹泻患者粪便样本的标准常规诊断算法很少适用于该病原体的检测，因此，除非专门检测，例如通过应用病原体特异性分子诊断方法，否则……很可能未被检测到。在本文介绍的研究中，我们在没有参考标准的测试比较中，使用来自加纳地方病流行区的高预测试概率粪便样本集，比较了三种针对……基因……（均为杂交探针检测法）和……（荧光共振能量转移检测法）的实时PCR检测方法。对1495份未显示PCR抑制迹象的粪便样本进行PCR检测，并应用潜在类别分析来评估实时PCR检测方法的诊断准确性。计算得出，……PCR检测的敏感性和特异性分别为93.0%和96.9%，……PCR检测为100%和98.2%，……PCR检测为12.7%和99.8%。在所评估的加纳人群中计算得出的……流行率为14.7%。如高滴度加标样本的检测结果所示，……检测法和……检测法与系统发育相关物种（如……）可能会发生交叉反应，但与系统发育距离较远的物种（如……）发生交叉反应的可能性较小。总之，……检测法表现出最有前景的性能特征，是唯一敏感性>95%的检测方法，尽管其95%置信区间较宽。此外，尽管已知该检测法与系统发育密切相关的物种（如……）存在交叉反应，但其特异性仍>98%，仍可接受。如果需要更高的确定性，对于……PCR检测结果为阳性的样本，可应用特异性接近100%的……检测法进行确认检测。然而，如果……检测法结果为阴性，由于……检测法灵敏度极低，不能可靠地排除在……检测法中检测到……的可能性。
