Universität zu Köln

Weinreich, Felix, Hahn, Andreas ORCID: 0000-0002-5157-8369, Eberhardt, Kirsten Alexandra, Feldt, Torsten, Sarfo, Fred Stephen, Di Cristanziano, Veronica, Frickmann, Hagen and Loderstaedt, Ulrike (2022). Comparison of Three Real-Time PCR Assays for the Detection of Cyclospora cayetanensis in Stool Samples Targeting the 18S rRNA Gene and the hsp70 Gene. Pathogens, 11 (2). BASEL: MDPI. ISSN 2076-0817

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Abstract

Diagnostic real-time PCR for the detection of Cyclospora cayetanensis in human stool samples has been applied for two decades. However, recent comparative assessments between in-house and commercial assays suggested room for improvement regarding the agreement of positive signals of the applied real-time PCRs. In order to assess the effect of the choice of the target sequence, 3 inhouse real time PCR assays targeting the 18S rRNA gene (n = 2, one of them later referred to as SSU rRNA gene assay to avoid confusion) and the hsp70 gene of C. cayetanensis were compared in a head-to-head comparison with 905 samples with high pretest probability for C. cayetanensis infections from Ghanaian HIV patients in a test comparison without a reference standard. Only slight agreement kappa of 0.095 was observed. In the assays targeting the SSU rRNA gene, the 18S rRNA gene, and hsp70, positive signals were recorded in 63, 45, and 0 instances, respectively, with latent class analysis-based estimation of sensitivity of 32.2%, 23.3%, 0% as well as of specificity of 99.7%, 99.9% and 100%, respectively. High cycle threshold values with an average of about 35 indicated low quantities of target DNA in the samples with similar Ct values in concordantly and discordantly positive samples. In conclusion, the study suggested target-gene-specific differences in the diagnostic accuracy of real-time PCR-based diagnosis of C. cayetanensis as well as an ongoing need for further standardization of this diagnostic approach.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Weinreich, Felix UNSPECIFIED UNSPECIFIED UNSPECIFIED Hahn, Andreas UNSPECIFIED orcid.org/0000-0002-5157-8369 UNSPECIFIED Eberhardt, Kirsten Alexandra UNSPECIFIED UNSPECIFIED UNSPECIFIED Feldt, Torsten UNSPECIFIED UNSPECIFIED UNSPECIFIED Sarfo, Fred Stephen UNSPECIFIED UNSPECIFIED UNSPECIFIED Di Cristanziano, Veronica UNSPECIFIED UNSPECIFIED UNSPECIFIED Frickmann, Hagen UNSPECIFIED UNSPECIFIED UNSPECIFIED Loderstaedt, Ulrike UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-689330
DOI:	10.3390/pathogens11020165
Journal or Publication Title:	Pathogens
Volume:	11
Number:	2
Date:	2022
Publisher:	MDPI
Place of Publication:	BASEL
ISSN:	2076-0817
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CRYPTOSPORIDIUM; CYSTOISOSPORA; DIAGNOSIS; BORNE Multiple languages Microbiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/68933