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 (2021). Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of Cryptosporidium spp. in Stool Samples. Pathogens, 10 (9). BASEL: MDPI. ISSN 2076-0817

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Abstract

As qualified microscopy of enteric parasitoses as defined by high diagnostic accuracy is difficult to maintain in non-endemic areas due to scarce opportunities for practicing with positive sample materials, molecular diagnostic options provide less investigator-dependent alternatives. Here, we compared three molecular targets for the real-time PCR-based detection of Cryptosporidium spp. From a population of 1000 individuals comprising both Ghanaian HIV (human immunodeficiency virus) patients and military returnees after deployment in the tropics, stool samples were assessed for Cryptosporidium spp. by real-time PCR targeting the small subunit ribosomal RNA (SSU rRNA) gene, the Cryptosporidium oocyst wall (COWP) gene, and the DnaJ-like protein gene (DnaJ), respectively. In declining order, sensitivity of 100% for the SSU rRNA gene PCR, 90.0% for the COWP PCR and 88.8% for the DnaJ PCR, respectively, as well as specificity of 99.6% for the COWP PCR and 96.9% for both the SSU rRNA gene PCR and the DnaJ PCR, respectively, were recorded. Substantial agreement (kappa value 0.663) between the three assays was observed. Further, an accuracy-adjusted Cryptosporidium spp. prevalence of 6.0% was calculated for the study population. In conclusion, none of the assessed real-time PCR assays were associated with perfect test accuracy. However, a combination of highly sensitive SSU rRNA gene PCR for screening purposes and more specific COWP PCR for confirmatory testing should allow reliable diagnosis of Cryptosporidium spp. in stool samples even in low prevalence settings.

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:38-598643
DOI:	10.3390/pathogens10091131
Journal or Publication Title:	Pathogens
Volume:	10
Number:	9
Date:	2021
Publisher:	MDPI
Place of Publication:	BASEL
ISSN:	2076-0817
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ENTAMOEBA-HISTOLYTICA; GIARDIA-LAMBLIA; INTESTINAL PROTOZOA; MULTIPLEX PCR; MOLECULAR DIAGNOSIS; PCR/RFLP ANALYSES; FECAL SAMPLES; PARVUM; PARASITES; QUANTIFICATION Multiple languages Microbiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/59864