Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum

Introduction: Rapid species identification and proper use of drug sare key requirements for the effective treatment and case management of tuberculosis. The development and evaluation of new diagnostic technique, which can diagnose causative agent in simple and rapid way, is the necessity of this ce...

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Main Author:	Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal
Format:	Technical Report
Language:	en_US
Published:	Nepal Health Research Council 2012
Subjects:	M. tuberculosis M avium complex M. kansasii LAMP
Online Access:	http://103.69.126.140:8080/handle/123456789/615
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spelling	oai:103.69.126.140:123456789-6152023-01-05T07:05:29Z Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal M. tuberculosis M avium complex M. kansasii LAMP Introduction: Rapid species identification and proper use of drug sare key requirements for the effective treatment and case management of tuberculosis. The development and evaluation of new diagnostic technique, which can diagnose causative agent in simple and rapid way, is the necessity of this century. Loop-Mediated Isothermal Amplification (LAMP) provides new possibilities of above requirements for direct detection of M. tuberculosis, M. avium complex and M. kansasiiin sputum samples. Objectives: To diagnose Mycobacaterium Tuberculosis early and accurately and to identify the prevalence of atypical mycobactera in Nepal. The study further intended to initiate LAMP as a diagnostic tools in hospital laboratories of Nepal. Materials and Methods:This study was carried out 130 sputum samples from suspected pulmonary tuberculosis patients were included in this study. All these samples were processed for flurochrome staining and were subjected to culture and LAMP. Thus sputum specimens were included in this study to compare them with microscopy, culture and LAMP. Results: Among them 50(38.46%) were found to bepositive by flurochrome staining, culture and LAMP. Similarly 48(36.92%) samples were negative by all diagnostic methods. 1(0.77%) microscopy and culture positive sample was negative by LAMP. Similarly 3(2.31%) microscopy and LAMP positive cases were negative by culture. 3(2.31%) culture positive cases were negative by both microscopy and LAMP. Eight (6.16%) culture negative cases were positive by LAMP where as 17(13.07%) microscopy negative samples were positive by culture and LAMP. Out of 78(100%) total LAMP positive cases, 76(97.44%) were positive for M. tuberculosis and remaining 2 (2.56%) were positive for M. intracellular. While comparing LAMP results with gold standard culture, the sensitivity, specificity, predictive value of positive test, predictive value of negative test, percentage of false negative and percentage of false positive of LAMP were found to be 94.36%, 81.36%, 85.90%, 92.31%, 5.63% and 18.64% respectively. Similarly, LAMP had sensitivity 98.14% and specificity 67.11% while compare with microscopy. Discussion and Conclusion: LAMP is highly sensitive and specific molecular technique, which can be used effectively for the diagnosis of clinically, microscopically, and culturally confusing cases thus facilitating the effective treatment and case management of tuberculosis and other atypical mycobacterial infection. Due to its easy operation and rapid amplification efficiency, it can be used in well-equipped laboratories for clinical use if sample preparation, nucleic acid extraction and cross-contamination controls are addressed. 2012-12-26T23:29:34Z 2022-11-08T10:17:07Z 2012-12-26T23:29:34Z 2022-11-08T10:17:07Z 2009 Technical Report http://103.69.126.140:8080/handle/123456789/615 en_US application/pdf Nepal Health Research Council
institution	My University
collection	DSpace
language	en_US
topic	M. tuberculosis M avium complex M. kansasii LAMP
spellingShingle	M. tuberculosis M avium complex M. kansasii LAMP Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
description	Introduction: Rapid species identification and proper use of drug sare key requirements for the effective treatment and case management of tuberculosis. The development and evaluation of new diagnostic technique, which can diagnose causative agent in simple and rapid way, is the necessity of this century. Loop-Mediated Isothermal Amplification (LAMP) provides new possibilities of above requirements for direct detection of M. tuberculosis, M. avium complex and M. kansasiiin sputum samples. Objectives: To diagnose Mycobacaterium Tuberculosis early and accurately and to identify the prevalence of atypical mycobactera in Nepal. The study further intended to initiate LAMP as a diagnostic tools in hospital laboratories of Nepal. Materials and Methods:This study was carried out 130 sputum samples from suspected pulmonary tuberculosis patients were included in this study. All these samples were processed for flurochrome staining and were subjected to culture and LAMP. Thus sputum specimens were included in this study to compare them with microscopy, culture and LAMP. Results: Among them 50(38.46%) were found to bepositive by flurochrome staining, culture and LAMP. Similarly 48(36.92%) samples were negative by all diagnostic methods. 1(0.77%) microscopy and culture positive sample was negative by LAMP. Similarly 3(2.31%) microscopy and LAMP positive cases were negative by culture. 3(2.31%) culture positive cases were negative by both microscopy and LAMP. Eight (6.16%) culture negative cases were positive by LAMP where as 17(13.07%) microscopy negative samples were positive by culture and LAMP. Out of 78(100%) total LAMP positive cases, 76(97.44%) were positive for M. tuberculosis and remaining 2 (2.56%) were positive for M. intracellular. While comparing LAMP results with gold standard culture, the sensitivity, specificity, predictive value of positive test, predictive value of negative test, percentage of false negative and percentage of false positive of LAMP were found to be 94.36%, 81.36%, 85.90%, 92.31%, 5.63% and 18.64% respectively. Similarly, LAMP had sensitivity 98.14% and specificity 67.11% while compare with microscopy. Discussion and Conclusion: LAMP is highly sensitive and specific molecular technique, which can be used effectively for the diagnosis of clinically, microscopically, and culturally confusing cases thus facilitating the effective treatment and case management of tuberculosis and other atypical mycobacterial infection. Due to its easy operation and rapid amplification efficiency, it can be used in well-equipped laboratories for clinical use if sample preparation, nucleic acid extraction and cross-contamination controls are addressed.
format	Technical Report
author	Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal
author_facet	Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal
author_sort	Nepal Health Research Council (NHRC), Ramshah Path, Kathmandu, Nepal
title	Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
title_short	Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
title_full	Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
title_fullStr	Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
title_full_unstemmed	Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum
title_sort	loop mediated isothermal amplification for the direct detection of human pulmonary infection with mycobacterium tuberculosis, mycobacterium avium complex and mycobacterium kansasii from sputum
publisher	Nepal Health Research Council
publishDate	2012
url	http://103.69.126.140:8080/handle/123456789/615
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_version_	1761501137527111680

Loop Mediated Isothermal Amplification For The Direct Detection of Human Pulmonary Infection with Mycobacterium Tuberculosis, Mycobacterium Avium Complex and Mycobacterium Kansasii from Sputum

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