Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples
<i>Cystoisospora (C.) belli</i> 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 ta...
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MDPI AG
2021-08-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/8/1053 |
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| author | Martin Blohm Andreas Hahn Ralf Matthias Hagen Kirsten Alexandra Eberhardt Holger Rohde Gérard Leboulle Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt |
| author_facet | Martin Blohm Andreas Hahn Ralf Matthias Hagen Kirsten Alexandra Eberhardt Holger Rohde Gérard Leboulle Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt |
| author_sort | Martin Blohm |
| collection | DOAJ |
| description | <i>Cystoisospora (C.) belli</i> 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 <i>C. belli</i> 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 <i>C. belli</i> 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. |
| first_indexed | 2024-03-10T08:29:11Z |
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| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T08:29:11Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-4e66484d596443629b7e3bfcd0dd5e082023-11-22T09:10:09ZengMDPI AGPathogens2076-08172021-08-01108105310.3390/pathogens10081053Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool SamplesMartin Blohm0Andreas Hahn1Ralf Matthias Hagen2Kirsten Alexandra Eberhardt3Holger Rohde4Gérard Leboulle5Torsten Feldt6Fred Stephen Sarfo7Veronica Di Cristanziano8Hagen Frickmann9Ulrike Loderstädt10Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, GermanyInstitute of Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, GermanyDepartment of Microbiology and Hospital Hygiene, Bundeswehr Central Hospital Koblenz, 56070 Koblenz, GermanyInstitute of Transfusion Medicine, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, GermanyInstitute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, GermanyTib MolBiol, Eresburgstraße 22-23, 12103 Berlin, GermanyDepartment of Gastroenterology, Hepatology and Infectious Diseases, University Medical Center Düsseldorf, 40225 Düsseldorf, GermanyDepartment of Medicine, Kwame Nkrumah University of Science and Technology, Kumasi 00233, GhanaInstitute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, 50935 Cologne, GermanyDepartment of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, GermanyDepartment of Hospital Hygiene & Infectious Diseases, University Medicine Göttingen, 37075 Göttingen, Germany<i>Cystoisospora (C.) belli</i> 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 <i>C. belli</i> 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 <i>C. belli</i> 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.https://www.mdpi.com/2076-0817/10/8/1053<i>Cystoisospora belli</i>diagnosisstoolreal-time PCRtest comparisonimmunosuppression |
| spellingShingle | Martin Blohm Andreas Hahn Ralf Matthias Hagen Kirsten Alexandra Eberhardt Holger Rohde Gérard Leboulle Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples Pathogens <i>Cystoisospora belli</i> diagnosis stool real-time PCR test comparison immunosuppression |
| title | Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples |
| title_full | Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples |
| title_fullStr | Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples |
| title_full_unstemmed | Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples |
| title_short | Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of <i>Cystoisospora belli</i> in Human Stool Samples |
| title_sort | comparison of two real time pcr assays targeting ribosomal sequences for the identification of i cystoisospora belli i in human stool samples |
| topic | <i>Cystoisospora belli</i> diagnosis stool real-time PCR test comparison immunosuppression |
| url | https://www.mdpi.com/2076-0817/10/8/1053 |
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