Cargando…

Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples

Cystoisospora (C.) belli is a coccidian parasite associated with acute or chronic gastroenteritis in immunocompromised patients. Dissatisfactory sensitivity of microscopy as the diagnostic standard approach has been described. Here, we comparatively evaluated two real-time PCRs targeting ribosomal R...

Descripción completa

Detalles Bibliográficos
Autores principales: Blohm, Martin, Hahn, Andreas, Hagen, Ralf Matthias, Eberhardt, Kirsten Alexandra, Rohde, Holger, Leboulle, Gérard, Feldt, Torsten, Sarfo, Fred Stephen, Di Cristanziano, Veronica, Frickmann, Hagen, Loderstädt, Ulrike
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399767/
https://www.ncbi.nlm.nih.gov/pubmed/34451517
http://dx.doi.org/10.3390/pathogens10081053
_version_ 1783745156259774464
author Blohm, Martin
Hahn, Andreas
Hagen, Ralf Matthias
Eberhardt, Kirsten Alexandra
Rohde, Holger
Leboulle, Gérard
Feldt, Torsten
Sarfo, Fred Stephen
Di Cristanziano, Veronica
Frickmann, Hagen
Loderstädt, Ulrike
author_facet Blohm, Martin
Hahn, Andreas
Hagen, Ralf Matthias
Eberhardt, Kirsten Alexandra
Rohde, Holger
Leboulle, Gérard
Feldt, Torsten
Sarfo, Fred Stephen
Di Cristanziano, Veronica
Frickmann, Hagen
Loderstädt, Ulrike
author_sort Blohm, Martin
collection PubMed
description Cystoisospora (C.) belli is a coccidian parasite associated with acute or chronic gastroenteritis in immunocompromised patients. Dissatisfactory sensitivity of microscopy as the diagnostic standard approach has been described. Here, we comparatively evaluated two real-time PCRs targeting ribosomal RNA gene sequences of C. belli in stool in a test comparison without a reference standard applying latent class analysis. Therefore, 1000 stool samples from Ghanaian HIV (human immunodeficiency virus) patients (n = 905) as well as military returnees from the tropics (n = 95) were assessed by both assays in parallel. After the exclusion of 33 samples showing PCR inhibition, 29 and 33 positive results were recorded with the 5.8S rRNA gene/ITS-2 sequence PCR and the ITS-2 sequence PCR, respectively, resulting in an accuracy-adjusted prevalence of 3.2%. Nearly perfect agreement between both assays was indicated by Fleiss’ kappa of 0.933 with sensitivity and specificity of 92.8% and 100% as well as 100% and 99.8% for the 5.8S rRNA gene/ITS-2 sequence PCR and the ITS-2 sequence PCR, respectively. Both assays proved to be suitable for the diagnosis of C. belli in human stool samples with slightly better sensitivity of the ITS-2 sequence assay, while the 5.8S rRNA gene/ITS-2 sequence PCR may be considered for confirmatory testing.
format Online
Article
Text
id pubmed-8399767
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-83997672021-08-29 Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples Blohm, Martin Hahn, Andreas Hagen, Ralf Matthias Eberhardt, Kirsten Alexandra Rohde, Holger Leboulle, Gérard Feldt, Torsten Sarfo, Fred Stephen Di Cristanziano, Veronica Frickmann, Hagen Loderstädt, Ulrike Pathogens Article Cystoisospora (C.) belli is a coccidian parasite associated with acute or chronic gastroenteritis in immunocompromised patients. Dissatisfactory sensitivity of microscopy as the diagnostic standard approach has been described. Here, we comparatively evaluated two real-time PCRs targeting ribosomal RNA gene sequences of C. belli in stool in a test comparison without a reference standard applying latent class analysis. Therefore, 1000 stool samples from Ghanaian HIV (human immunodeficiency virus) patients (n = 905) as well as military returnees from the tropics (n = 95) were assessed by both assays in parallel. After the exclusion of 33 samples showing PCR inhibition, 29 and 33 positive results were recorded with the 5.8S rRNA gene/ITS-2 sequence PCR and the ITS-2 sequence PCR, respectively, resulting in an accuracy-adjusted prevalence of 3.2%. Nearly perfect agreement between both assays was indicated by Fleiss’ kappa of 0.933 with sensitivity and specificity of 92.8% and 100% as well as 100% and 99.8% for the 5.8S rRNA gene/ITS-2 sequence PCR and the ITS-2 sequence PCR, respectively. Both assays proved to be suitable for the diagnosis of C. belli in human stool samples with slightly better sensitivity of the ITS-2 sequence assay, while the 5.8S rRNA gene/ITS-2 sequence PCR may be considered for confirmatory testing. MDPI 2021-08-19 /pmc/articles/PMC8399767/ /pubmed/34451517 http://dx.doi.org/10.3390/pathogens10081053 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Blohm, Martin
Hahn, Andreas
Hagen, Ralf Matthias
Eberhardt, Kirsten Alexandra
Rohde, Holger
Leboulle, Gérard
Feldt, Torsten
Sarfo, Fred Stephen
Di Cristanziano, Veronica
Frickmann, Hagen
Loderstädt, Ulrike
Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title_full Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title_fullStr Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title_full_unstemmed Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title_short Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples
title_sort comparison of two real-time pcr assays targeting ribosomal sequences for the identification of cystoisospora belli in human stool samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399767/
https://www.ncbi.nlm.nih.gov/pubmed/34451517
http://dx.doi.org/10.3390/pathogens10081053
work_keys_str_mv AT blohmmartin comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT hahnandreas comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT hagenralfmatthias comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT eberhardtkirstenalexandra comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT rohdeholger comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT leboullegerard comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT feldttorsten comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT sarfofredstephen comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT dicristanzianoveronica comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT frickmannhagen comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples
AT loderstadtulrike comparisonoftworealtimepcrassaystargetingribosomalsequencesfortheidentificationofcystoisosporabelliinhumanstoolsamples