Global Advanced Research Journal of Medicine and Medical Sciences (GARJMMS) ISSN: 2315-5159
April 2018, Vol. 7(4), pp. 085-089
Copyright © 2018 Global Advanced Research Journals

 

Full Length Research Article

Diagnostic accuracy study of loop mediated isothermal amplification in the detection of Mycobacterium tuberculosis in Pakistan

Asim Saeed*, Afreenish Hassan, Umme Farwa, Shahid Ahmad Abbasi and Asim Ali Shah

Microbiologist National Institute of Health, Islamabad, Pakistan, Senior Pathologist/Microbiologist Excel Labs (Pvt) Ltd

*Corresponding Author E-mail: dr.asimch@gmail.com; Cell: +923145183024

Accepted 20 April, 2018 

Abstract

The study has been conducted in Armed Forces Institute of Pathology, Rawalpindi Pakistan from 2015 to 2017. The study was done to determine the adequacy of Loop-mediated Isothermal Amplification for the detection of Mycobacterium tuberculosis. A cross sectional study was done on 72 clinical sputum samples taken from the patient suspected of tuberculosis using a novel technique LAMP targeting IS6110 gene sequence for the first time in Pakistan. Six primers recognizing eight distinct regions on the target sequence were employed. The assay was performed in total volume of 25 μl containing primers, DNA polymerase, Fluorescent dye and sample incubated at 65°C for 60 minutes along with positive and negative controls. LAMP amplicons were detected by their fluorescence under UV light and compared with gold standard MGIT culture system and smear microscopy. Out of 72 total samples 60(83.3%) were LAMP positive and 61(84.7%) were MGIT positive. Its specificity, sensitivity is 100% and 98.36% respectively with PPV 100% and NPV 91.6%. According to our study the LAMP assay is suggested to be a potential nucleic acid based diagnostic method for TB detection in developing countries.

Keywords: Loop Mediated Isothermal Amplification (LAMP), IS6110 gene, Mycobacterium tuberculosis.   Mycobacterium Growth Indicator Tube (MGIT)

 

References

Aryan E, Makvandi M, Farajzadeh A, Huygen K, Bifani P, Mousavi SL, Romano M (2010). A novel and more sensitive loop-mediated isothermal amplification assay targeting IS6110 for detection of Mycobacterium tuberculosis complex. Microbiol. Res. 165(3): 211-220.

Bi A, Nakajima C, Fukushima Y, Tamaru A, Sugawara I, Kimura A, Suzuki Y (2012). A rapid loop-mediated isothermal amplification assay targeting hspX for the detection of Mycobacterium tuberculosis complex. Jap. J. Infect. Dis. 65(3): 247-251.

Boehme CC, Nabeta P, Henostroza G, Raqib R, Rahim Z, Gerhardt M, Hase T (2007). Operational feasibility of using loop-mediated isothermal amplification for diagnosis of pulmonary tuberculosis in microscopy centers of developing countries. J. Clin. Microbiol. 45(6): 1936-1940.

Desikan P (2013). Sputum smear microscopy in tuberculosis: Is it still relevant? The Ind. J. Med. Res 137(3): 442.

Ganeswrie R, Chui C, Balan S, Puthucheary S (2004). Comparison of BACTEC MGIT 960 system and BACTEC 460 TB system for growth and detection of Mycobacteria from clinical specimens. The Malaysian J. Pathol. 26(2): 99-103.

Geojith G, Dhanasekaran S, Chandran SP, Kenneth J (2011). Efficacy of loop mediated isothermal amplification (LAMP) assay for the laboratory identification of Mycobacterium tuberculosis isolates in a resource limited setting. J. Microbiol. Methods. 84(1): 71-73.

Iwamoto T, Sonobe T, Hayashi K (2003). Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, M. avium, and M. intracellulare in sputum samples. J. Clin. Microbiol.41(6): 2616-2622.

Khan MB, Mashhood AA, Qureishi SM (2016). Recent advances in the management of tuberculosis. J. Pak. Assoc. Dermatol. 15(2): 151-160.

Kohan L, Shahhosseiny MH, Razavi MR, Parivar K, Moslemi E, Werngren J (2011). Evaluation of loop mediated isothermal amplification for diagnosis of Mycobacterium tuberculosis complex in clinical samples. Afr. J. Biotechnol. 10(26): 5096-5101.

Organization WH (2013). Global tuberculosis report 2013: World Health Organization.

Palomino J, Portaels F (1998). Effects of decontamination methods and culture conditions on viability of Mycobacterium ulcerans in the BACTEC system. J. Clin. Microbiol.36(2): 402-408.

Pandey BD, Poudel A, Yoda T, Tamaru A, Oda N, Fukushima Y, Sapkota B (2008). Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients. J. Med. Microbiol. 57(4), 439-443.

Ryu YJ (2015). Diagnosis of pulmonary tuberculosis: recent advances and diagnostic algorithms. Tuberculosis and respiratory diseases. 78(2): 64-71.

Vermund SH, Altaf A, Samo RN, Khanani R, Baloch N, Qadeer E, Shah SA (2009). Tuberculosis in Pakistan: A decade of progress, a future of challenge. J. Pak. Med. Assoc. 59(4): 1-8.

Watterson S, Drobniewski F (2000). Modern laboratory diagnosis of mycobacterial infections. J. Clin. Pathol. 53(10): 727-732.

Zhu RY, Zhang KX, Zhao MQ, Liu YH, Xu YY, Ju CM, Chen JD (2009). Use of visual loop-mediated isotheral amplification of rimM sequence for rapid detection of Mycobacterium tuberculosis and Mycobacterium bovis. J. Microbiol. Methods. 78(3): 339-343.


Current Issue

Viewing Options

View Full Article - PDF
Download Full Article - PDF

Search for Articles

Asim Saeed on Google Scholar
Asim Saeed on Pubmed
Afreenish Hassan on Google Scholar
Afreenish Hassan on Pubmed
Umme Farwa on Google Scholar
Umme Farwa on Pubmed
Shahid Ahmad Abbasi on Google Scholar
Shahid Ahmad Abbasi on Pubmed
Asim Ali Shah on Google Scholar
Asim Ali Shah on Pubmed

Statistics

Viewed 212
Printed 70
Downloaded 148
Powered By iPortal Works