Zautner Andreas Erich, Frickmann Hagen, Hahn Andreas, Sarfo Fred Stephen, Norman Betty Roberta, Dompreh Albert, Agyei Martin Kofi, Asibey Shadrack Osei, Boateng Richard, Kuffour Edmund Osei, Di Cristanziano Veronica, Tufa Tafese Beyene, Feldt Torsten, Eberhardt Kirsten Alexandra
Institute of Medical Microbiology and Hospital Hygiene, Medical Faculty, Otto-von-Guericke University Magdeburg, 39120 Magdeburg, Germany.
Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 22049 Hamburg, Germany.
Microorganisms. 2025 Aug 24;13(9):1976. doi: 10.3390/microorganisms13091976.
Molecular diagnoses of and in human samples are becoming increasingly common. To contribute to the ongoing standardization of molecular diagnostic approaches targeting these parasites, we compared three published - and -specific real-time PCR assays in test comparisons without a reference standard. Latent class analysis (LCA) was used to calculate diagnostic accuracy estimations for the three compared assays per parameter. The comparison was conducted using stool samples from Ghanaian individuals. In the course of the assessment of 873 stool samples, the number of detected positive PCR results ranged from 10 to 15 for and from 4 to 54 for depending on the applied assay. Diagnostic accuracy estimates of real-time PCR sensitivity for and ranged from 89% to 100% and from 75% to 100%, respectively; diagnostic estimates of specificity ranged from 99% to 100% and from 94% to 100%, respectively. Diagnostic accuracy-adjusted prevalence estimates were 1.2% for and 0.5% for . High cycle threshold values of real-time PCR > 35 showed a particularly reduced likeliness of reproducibility when applying competitor real-time PCR assays. There were no clear-cut differences in terms of diagnostic accuracy favoring either small-subunit ribosomal ribonucleic acid (SSU rRNA) gene sequences or the dispersed repetitive sequence for PCR. The same applied to the comparison of real-time PCRs targeting SSU rRNA gene sequences and the SSU rRNA episomal repeat sequence (SREPH) of . In conclusion, interchangeability of the compared real-time PCR assays was higher for the assessed assays compared with the assessed assays. Regional diagnostic accuracy testing seems advisable before literature-adapted assays for rare tropical pathogens like and are applied in different study regions.
对人体样本中的[寄生虫名称1]和[寄生虫名称2]进行分子诊断正变得越来越普遍。为推动针对这些寄生虫的分子诊断方法的持续标准化，我们在没有参考标准的测试比较中，比较了三种已发表的针对[寄生虫名称1]和[寄生虫名称2]的特异性实时荧光定量PCR检测方法。使用潜在类别分析（LCA）来计算每个参数下三种比较检测方法的诊断准确性估计值。比较是使用来自加纳个体的粪便样本进行的。在对873份粪便样本的评估过程中，根据所应用的检测方法，检测到的[寄生虫名称1]阳性PCR结果数量在10至15之间波动，而[寄生虫名称2]的阳性结果数量则在4至54之间波动。针对[寄生虫名称1]和[寄生虫名称2]的实时荧光定量PCR敏感性的诊断准确性估计值分别为89%至100%和75%至100%；特异性的诊断估计值分别为99%至100%和94%至100%。经诊断准确性调整后的患病率估计值，[寄生虫名称1]为1.2%，[寄生虫名称2]为0.5%。当应用竞争实时荧光定量PCR检测方法时，实时荧光定量PCR > 35的高循环阈值显示出特别低的重现性可能性。在诊断准确性方面，对于[寄生虫名称1]的PCR，无论是小亚基核糖体核糖核酸（SSU rRNA）基因序列还是[特定重复序列名称][寄生虫名称1]分散重复序列，都没有明显的差异。对于针对[寄生虫名称2]的SSU rRNA基因序列和SSU rRNA附加体重复序列（SREPH）的实时荧光定量PCR比较也是如此。总之，与评估的[寄生虫名称2]检测方法相比，评估的[寄生虫名称1]检测方法中，所比较的实时荧光定量PCR检测方法的互换性更高。在针对[寄生虫名称1]和[寄生虫名称2]等罕见热带病原体的文献适配检测方法应用于不同研究区域之前，进行区域诊断准确性测试似乎是可取的。
