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Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients

Pneumocystis pneumonia (PCP) is one of the common opportunistic infection among HIV and non-HIV immunocompromised patients. The lack of a rapid and specific diagnostic test necessitates a more reliable laboratory diagnostic test for PCP. In the present study, the loop-mediated isothermal amplificati...

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Autores principales: Singh, Preeti, Singh, Sundeep, Mirdha, Bijay Ranjan, Guleria, Randeep, Agarwal, Sanjay Kumar, Mohan, Anant
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668309/
https://www.ncbi.nlm.nih.gov/pubmed/26664746
http://dx.doi.org/10.1155/2015/819091
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author Singh, Preeti
Singh, Sundeep
Mirdha, Bijay Ranjan
Guleria, Randeep
Agarwal, Sanjay Kumar
Mohan, Anant
author_facet Singh, Preeti
Singh, Sundeep
Mirdha, Bijay Ranjan
Guleria, Randeep
Agarwal, Sanjay Kumar
Mohan, Anant
author_sort Singh, Preeti
collection PubMed
description Pneumocystis pneumonia (PCP) is one of the common opportunistic infection among HIV and non-HIV immunocompromised patients. The lack of a rapid and specific diagnostic test necessitates a more reliable laboratory diagnostic test for PCP. In the present study, the loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of Pneumocystis jirovecii. 185 clinical respiratory samples, including both BALF and IS, were subjected to GMS staining, nested PCR, and LAMP assay. Of 185 respiratory samples, 12/185 (6.5%), 41/185 (22.2%), and 49/185 (26.5%) samples were positive by GMS staining, nested PCR, and LAMP assay, respectively. As compared to nested PCR, additional 8 samples were positive by LAMP assay and found to be statistically significant (p < 0.05) with the detection limit of 1 pg. Thus, the LAMP assay may serve as a better diagnostic tool for the detection of P. jirovecii with high sensitivity and specificity, less turn-around time, operational simplicity, single-step amplification, and immediate visual detection.
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spelling pubmed-46683092015-12-10 Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients Singh, Preeti Singh, Sundeep Mirdha, Bijay Ranjan Guleria, Randeep Agarwal, Sanjay Kumar Mohan, Anant Mol Biol Int Research Article Pneumocystis pneumonia (PCP) is one of the common opportunistic infection among HIV and non-HIV immunocompromised patients. The lack of a rapid and specific diagnostic test necessitates a more reliable laboratory diagnostic test for PCP. In the present study, the loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of Pneumocystis jirovecii. 185 clinical respiratory samples, including both BALF and IS, were subjected to GMS staining, nested PCR, and LAMP assay. Of 185 respiratory samples, 12/185 (6.5%), 41/185 (22.2%), and 49/185 (26.5%) samples were positive by GMS staining, nested PCR, and LAMP assay, respectively. As compared to nested PCR, additional 8 samples were positive by LAMP assay and found to be statistically significant (p < 0.05) with the detection limit of 1 pg. Thus, the LAMP assay may serve as a better diagnostic tool for the detection of P. jirovecii with high sensitivity and specificity, less turn-around time, operational simplicity, single-step amplification, and immediate visual detection. Hindawi Publishing Corporation 2015 2015-11-19 /pmc/articles/PMC4668309/ /pubmed/26664746 http://dx.doi.org/10.1155/2015/819091 Text en Copyright © 2015 Preeti Singh et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Singh, Preeti
Singh, Sundeep
Mirdha, Bijay Ranjan
Guleria, Randeep
Agarwal, Sanjay Kumar
Mohan, Anant
Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title_full Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title_fullStr Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title_full_unstemmed Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title_short Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients
title_sort evaluation of loop-mediated isothermal amplification assay for the detection of pneumocystis jirovecii in immunocompromised patients
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668309/
https://www.ncbi.nlm.nih.gov/pubmed/26664746
http://dx.doi.org/10.1155/2015/819091
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