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...
Autores principales: | , , , , , , , , , , |
---|---|
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 |