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Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates

INTRODUCTION: Pseudomonas aeruginosa is a common cause of ventilator-associated pneumonia (VAP). Rapid and accurate detection of lower respiratory tract colonization and/or infection with P. aeruginosa may advise targeted preventive (antibody-based) strategies and antibiotic therapy. To investigate...

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Autores principales: van der Schalk, Thomas Ewout, Coppens, Jasmine, Timbermont, Leen, Turlej-Rogacka, Agata, Van Heirstraeten , Liesbet, Berkell, Matilda, Yu, Li, Lammens, Christine, Xavier, Basil Britto, Matheeussen, Veerle, Ieven, Margareta, McCarthy, Michael, Jorens, Philippe G., Ruzin, Alexey, Esser, Mark T., Kumar-Singh, Samir, Goossens, Herman, Malhotra-Kumar, Surbhi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8300976/
https://www.ncbi.nlm.nih.gov/pubmed/34301343
http://dx.doi.org/10.1186/s13756-021-00978-9
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author van der Schalk, Thomas Ewout
Coppens, Jasmine
Timbermont, Leen
Turlej-Rogacka, Agata
Van Heirstraeten , Liesbet
Berkell, Matilda
Yu, Li
Lammens, Christine
Xavier, Basil Britto
Matheeussen, Veerle
Ieven, Margareta
McCarthy, Michael
Jorens, Philippe G.
Ruzin, Alexey
Esser, Mark T.
Kumar-Singh, Samir
Goossens, Herman
Malhotra-Kumar, Surbhi
author_facet van der Schalk, Thomas Ewout
Coppens, Jasmine
Timbermont, Leen
Turlej-Rogacka, Agata
Van Heirstraeten , Liesbet
Berkell, Matilda
Yu, Li
Lammens, Christine
Xavier, Basil Britto
Matheeussen, Veerle
Ieven, Margareta
McCarthy, Michael
Jorens, Philippe G.
Ruzin, Alexey
Esser, Mark T.
Kumar-Singh, Samir
Goossens, Herman
Malhotra-Kumar, Surbhi
author_sort van der Schalk, Thomas Ewout
collection PubMed
description INTRODUCTION: Pseudomonas aeruginosa is a common cause of ventilator-associated pneumonia (VAP). Rapid and accurate detection of lower respiratory tract colonization and/or infection with P. aeruginosa may advise targeted preventive (antibody-based) strategies and antibiotic therapy. To investigate this, we compared semi-quantitative culture results from 80 endotracheal aspirates (ETA) collected from mechanically-ventilated patients, to two culture and two non-culture-based methods for detection of P. aeruginosa. METHODS: P. aeruginosa-positive (n = 40) and -negative (n = 40) ETAs from mechanically ventilated patients analyzed initally by (i) routine semi-quantitative culture, were further analyzed with (ii) quantitative culture on chromogenic ChromID P. aeruginosa and blood agar; (iii) enrichment in brain heart infusion broth followed by plating on blood agar and ChromID P. aeruginosa; (iv) O-antigen acetylase gene-based TaqMan qPCR; and (v) GeneXpert PA PCR assay. RESULTS: Of the 80 ETA samples included, one sample that was negative for P. aeruginosa by semi-quantitative culture was found to be positive by the other four methods, and was included in an “extended” gold standard panel. Based on this extended gold standard, both semi-quantitative culture and the GeneXpert PA assay showed 97.6% sensitivity and 100% specificity. The quantitative culture, enrichment culture and O-antigen acetylase gene-based TaqMan qPCR had a sensitivity of 97.6%, 89.5%, 92.7%, and a specificity of 97.4%, 100%, and 71.1%, respectively. CONCLUSION: This first evaluation of the GeneXpert PA assay with ETA samples found it to be as sensitive and specific as the routine, hospital-based semi-quantitative culture method. Additionally, the GeneXpert PA assay is easy to perform (hands-on time ≈ 5 min) and rapid (≈ 55 min assay time). The combination of the high sensitivity and high specificity together with the rapid acquisition of results makes the GeneXpert PA assay a highly recommended screening technique. Where this equipment is not available, semi-quantitative culture remains the most sensitive of the culture methods evaluated here for P. aeruginosa detection in ETA samples. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13756-021-00978-9.
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spelling pubmed-83009762021-07-26 Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates van der Schalk, Thomas Ewout Coppens, Jasmine Timbermont, Leen Turlej-Rogacka, Agata Van Heirstraeten , Liesbet Berkell, Matilda Yu, Li Lammens, Christine Xavier, Basil Britto Matheeussen, Veerle Ieven, Margareta McCarthy, Michael Jorens, Philippe G. Ruzin, Alexey Esser, Mark T. Kumar-Singh, Samir Goossens, Herman Malhotra-Kumar, Surbhi Antimicrob Resist Infect Control Research INTRODUCTION: Pseudomonas aeruginosa is a common cause of ventilator-associated pneumonia (VAP). Rapid and accurate detection of lower respiratory tract colonization and/or infection with P. aeruginosa may advise targeted preventive (antibody-based) strategies and antibiotic therapy. To investigate this, we compared semi-quantitative culture results from 80 endotracheal aspirates (ETA) collected from mechanically-ventilated patients, to two culture and two non-culture-based methods for detection of P. aeruginosa. METHODS: P. aeruginosa-positive (n = 40) and -negative (n = 40) ETAs from mechanically ventilated patients analyzed initally by (i) routine semi-quantitative culture, were further analyzed with (ii) quantitative culture on chromogenic ChromID P. aeruginosa and blood agar; (iii) enrichment in brain heart infusion broth followed by plating on blood agar and ChromID P. aeruginosa; (iv) O-antigen acetylase gene-based TaqMan qPCR; and (v) GeneXpert PA PCR assay. RESULTS: Of the 80 ETA samples included, one sample that was negative for P. aeruginosa by semi-quantitative culture was found to be positive by the other four methods, and was included in an “extended” gold standard panel. Based on this extended gold standard, both semi-quantitative culture and the GeneXpert PA assay showed 97.6% sensitivity and 100% specificity. The quantitative culture, enrichment culture and O-antigen acetylase gene-based TaqMan qPCR had a sensitivity of 97.6%, 89.5%, 92.7%, and a specificity of 97.4%, 100%, and 71.1%, respectively. CONCLUSION: This first evaluation of the GeneXpert PA assay with ETA samples found it to be as sensitive and specific as the routine, hospital-based semi-quantitative culture method. Additionally, the GeneXpert PA assay is easy to perform (hands-on time ≈ 5 min) and rapid (≈ 55 min assay time). The combination of the high sensitivity and high specificity together with the rapid acquisition of results makes the GeneXpert PA assay a highly recommended screening technique. Where this equipment is not available, semi-quantitative culture remains the most sensitive of the culture methods evaluated here for P. aeruginosa detection in ETA samples. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13756-021-00978-9. BioMed Central 2021-07-23 /pmc/articles/PMC8300976/ /pubmed/34301343 http://dx.doi.org/10.1186/s13756-021-00978-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
van der Schalk, Thomas Ewout
Coppens, Jasmine
Timbermont, Leen
Turlej-Rogacka, Agata
Van Heirstraeten , Liesbet
Berkell, Matilda
Yu, Li
Lammens, Christine
Xavier, Basil Britto
Matheeussen, Veerle
Ieven, Margareta
McCarthy, Michael
Jorens, Philippe G.
Ruzin, Alexey
Esser, Mark T.
Kumar-Singh, Samir
Goossens, Herman
Malhotra-Kumar, Surbhi
Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title_full Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title_fullStr Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title_full_unstemmed Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title_short Evaluation of GeneXpert PA assay compared to genomic and (semi-)quantitative culture methods for direct detection of Pseudomonas aeruginosa in endotracheal aspirates
title_sort evaluation of genexpert pa assay compared to genomic and (semi-)quantitative culture methods for direct detection of pseudomonas aeruginosa in endotracheal aspirates
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8300976/
https://www.ncbi.nlm.nih.gov/pubmed/34301343
http://dx.doi.org/10.1186/s13756-021-00978-9
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