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

Standard

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; Cristanziano, Veronica Di; Frickmann, Hagen; Loderstädt, Ulrike.

In: PATHOGENS, Vol. 10, No. 8, 19.08.2021.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Blohm, M, Hahn, A, Hagen, RM, Eberhardt, KA, Rohde, H, Leboulle, G, Feldt, T, Sarfo, FS, Cristanziano, VD, Frickmann, H & Loderstädt, U 2021, 'Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples', PATHOGENS, vol. 10, no. 8. https://doi.org/10.3390/pathogens10081053

APA

Blohm, M., Hahn, A., Hagen, R. M., Eberhardt, K. A., Rohde, H., Leboulle, G., Feldt, T., Sarfo, F. S., Cristanziano, V. D., Frickmann, H., & Loderstädt, U. (2021). Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples. PATHOGENS, 10(8). https://doi.org/10.3390/pathogens10081053

Vancouver

Blohm M, Hahn A, Hagen RM, Eberhardt KA, Rohde H, Leboulle G et al. Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples. PATHOGENS. 2021 Aug 19;10(8). https://doi.org/10.3390/pathogens10081053

Bibtex

@article{bcaa1ba0f5f74b7ca1d653c041465f9d,
title = "Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples",
abstract = "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{\textquoteright} 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",
author = "Martin Blohm and Andreas Hahn and Hagen, {Ralf Matthias} and Eberhardt, {Kirsten Alexandra} and Holger Rohde and G{\'e}rard Leboulle and Torsten Feldt and Sarfo, {Fred Stephen} and Cristanziano, {Veronica Di} and Hagen Frickmann and Ulrike Loderst{\"a}dt",
year = "2021",
month = aug,
day = "19",
doi = "10.3390/pathogens10081053",
language = "English",
volume = "10",
journal = "PATHOGENS",
issn = "2076-0817",
publisher = "MDPI AG",
number = "8",

}

RIS

TY - JOUR

T1 - Comparison of Two Real-Time PCR Assays Targeting Ribosomal Sequences for the Identification of Cystoisospora belli in Human Stool Samples

AU - Blohm, Martin

AU - Hahn, Andreas

AU - Hagen, Ralf Matthias

AU - Eberhardt, Kirsten Alexandra

AU - Rohde, Holger

AU - Leboulle, Gérard

AU - Feldt, Torsten

AU - Sarfo, Fred Stephen

AU - Cristanziano, Veronica Di

AU - Frickmann, Hagen

AU - Loderstädt, Ulrike

PY - 2021/8/19

Y1 - 2021/8/19

N2 - 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

AB - 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

UR - https://doi.org/10.3390/pathogens10081053

U2 - 10.3390/pathogens10081053

DO - 10.3390/pathogens10081053

M3 - SCORING: Journal article

VL - 10

JO - PATHOGENS

JF - PATHOGENS

SN - 2076-0817

IS - 8

ER -