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Normalized real-time PCR for diagnosis of H. pylori infection

Objectives: There is increasing interest in the use of quantitative PCR (q-PCR) for diagnosis of H. pylori infection. However, the assay remains largely unstandardized, making comparison between studies unreliable. The objective of this study was to assess accuracy of a normalized q-PCR assay for di...

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Autores principales: Al-Moayad, Ebtisam E., Alghalibi, Saeed M., Al-Shamahy, Hassan A., Nasher, Akram T., Al-hebshi, Nezar N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bloomsbury Qatar Foundation Journals 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4344986/
https://www.ncbi.nlm.nih.gov/pubmed/25745602
http://dx.doi.org/10.5339/qmj.2014.19
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author Al-Moayad, Ebtisam E.
Alghalibi, Saeed M.
Al-Shamahy, Hassan A.
Nasher, Akram T.
Al-hebshi, Nezar N.
author_facet Al-Moayad, Ebtisam E.
Alghalibi, Saeed M.
Al-Shamahy, Hassan A.
Nasher, Akram T.
Al-hebshi, Nezar N.
author_sort Al-Moayad, Ebtisam E.
collection PubMed
description Objectives: There is increasing interest in the use of quantitative PCR (q-PCR) for diagnosis of H. pylori infection. However, the assay remains largely unstandardized, making comparison between studies unreliable. The objective of this study was to assess accuracy of a normalized q-PCR assay for diagnosis of the infection. Subjects and methods: Seventy-six fresh gastric biopsy specimens were collected from patients undergoing upper gastrointestinal tract endoscopy and examined by rapid urease test (RUT), culture, and a commercial TaqMan q-PCR assay targeting the ureA gene. Counts obtained from the latter assay were normalized to the human ACTB gene. A subject was considered to be infected if two or more assays were positive. Results: The detection rates were 42.1%, 52.6%, and 78.9% by culture, RUT and q-PCR, respectively. Bacterial density ranged 0.005 to 4800 bacteria per 100 human cells. Because q-PCR showed low initial specificity (45.7%), the cutoff value for the assay was recalculated as 1 bacterium per 100 human cells, using ROC curve analysis. Accordingly, the sensitivities and specificities were 79.5% and 97.3%, respectively, for culture; 94.9% and 91.9%, respectively, for RUT; and 94.9% and 94.6%, respectively, for q-PCR. By gold standard, 39 of the dyspeptic patients (51.3%) were found to be infected. Conclusions: With the identified cutoff value, the q-PCR assay diagnosed H. pylori infection with an accuracy slightly superior to that of RUT. However, the possibility that low counts detected only by q-PCR represent true infections warrants further investigation. Normalization of bacterial counts for standardization of q-PCR H. pylori assays is recommended.
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spelling pubmed-43449862015-03-05 Normalized real-time PCR for diagnosis of H. pylori infection Al-Moayad, Ebtisam E. Alghalibi, Saeed M. Al-Shamahy, Hassan A. Nasher, Akram T. Al-hebshi, Nezar N. Qatar Med J Research Article Objectives: There is increasing interest in the use of quantitative PCR (q-PCR) for diagnosis of H. pylori infection. However, the assay remains largely unstandardized, making comparison between studies unreliable. The objective of this study was to assess accuracy of a normalized q-PCR assay for diagnosis of the infection. Subjects and methods: Seventy-six fresh gastric biopsy specimens were collected from patients undergoing upper gastrointestinal tract endoscopy and examined by rapid urease test (RUT), culture, and a commercial TaqMan q-PCR assay targeting the ureA gene. Counts obtained from the latter assay were normalized to the human ACTB gene. A subject was considered to be infected if two or more assays were positive. Results: The detection rates were 42.1%, 52.6%, and 78.9% by culture, RUT and q-PCR, respectively. Bacterial density ranged 0.005 to 4800 bacteria per 100 human cells. Because q-PCR showed low initial specificity (45.7%), the cutoff value for the assay was recalculated as 1 bacterium per 100 human cells, using ROC curve analysis. Accordingly, the sensitivities and specificities were 79.5% and 97.3%, respectively, for culture; 94.9% and 91.9%, respectively, for RUT; and 94.9% and 94.6%, respectively, for q-PCR. By gold standard, 39 of the dyspeptic patients (51.3%) were found to be infected. Conclusions: With the identified cutoff value, the q-PCR assay diagnosed H. pylori infection with an accuracy slightly superior to that of RUT. However, the possibility that low counts detected only by q-PCR represent true infections warrants further investigation. Normalization of bacterial counts for standardization of q-PCR H. pylori assays is recommended. Bloomsbury Qatar Foundation Journals 2014-12-09 /pmc/articles/PMC4344986/ /pubmed/25745602 http://dx.doi.org/10.5339/qmj.2014.19 Text en © 2014 Al-Moayad, Alghalibi, Al-Shamahy, Nasher, Al-hebshi, licensee Bloomsbury Qatar Foundation Journals. This is an open access article distributed under the terms of the Creative Commons Attribution license CC BY 4.0, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Al-Moayad, Ebtisam E.
Alghalibi, Saeed M.
Al-Shamahy, Hassan A.
Nasher, Akram T.
Al-hebshi, Nezar N.
Normalized real-time PCR for diagnosis of H. pylori infection
title Normalized real-time PCR for diagnosis of H. pylori infection
title_full Normalized real-time PCR for diagnosis of H. pylori infection
title_fullStr Normalized real-time PCR for diagnosis of H. pylori infection
title_full_unstemmed Normalized real-time PCR for diagnosis of H. pylori infection
title_short Normalized real-time PCR for diagnosis of H. pylori infection
title_sort normalized real-time pcr for diagnosis of h. pylori infection
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4344986/
https://www.ncbi.nlm.nih.gov/pubmed/25745602
http://dx.doi.org/10.5339/qmj.2014.19
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