UTILITY OF ANTIGEN DETECTION TEST AND POLYMERASE CHAIN REACTION IN THE DIFFERENTIATION OF TUBERCULOUS AND NON-TUBERCULOUS MYCOBACTERIA
DOI:
https://doi.org/10.22159/ajpcr.2017.v10i6.18261Keywords:
Mycobacterium tuberculosis, Polymerase chain reaction, Culture, Antigen detectionAbstract
Objectives: Cultivation and identification of mycobacteria to species level remains difficult and time-consuming. Hence, easy and rapid diagnostic methods are necessary for the differentiation of Mycobacterium tuberculosis (MTB) from non-tuberculous mycobacteria (NTM). The present study aims to detect and differentiate MTB from NTM isolated from clinical samples by immunochromatographic test (ICT) and polymerase chain reaction (PCR).
Methods: Over a period of 1 year, clinical samples (n=496) received from suspected cases of TB, at the Department of Microbiology, Kasturba Medical College Hospital, Mangalore were cultured to isolate Mycobacterium spp. Identification of all the isolates was done by conventional biochemical technique, ICT, and PCR.
Results: Among the 496 samples processed, 49 (9.87%) were acid-fast bacilli smear positive and 59 (11.89%) samples showed the growth of Mycobacterium spp. Among these, 10 were rapid growers, 49 were slow-growing mycobacteria, out of which 30 were MTB as identified by conventional biochemical reaction. Out of 59 Mycobacterial isolates subjected to ICT for the detection of MPT 64 antigen, only 28 were identified as MTB. However, all the 30 isolates were correctly identified as MTB by PCR.
Conclusion: Hence, PCR is essential for rapid differentiation of non-tuberculous Mycobacterium from MTB. False negative results seen with immunochromatographic MPT 64 antigen assay could be due to mutations within the mpt64 gene. Further studies are necessary to characterize these PCR-positive and immunochromatographic assay negative MTB isolates.
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References
TB India. Revised National Tuberculosis Control Programme. Annual Status Report. Central TB Division, Directorate General of Health Services, Ministry of Health and Family Welfare, Nirman Bhavan, New Delhi; 2014. Available from: http://www.tbcindia.nic.in/index1. php?lang=1&level=1&sublinkid=4160&lid=2807. [Last accessed on 2016 Jan 26].
Global Tuberculosis Report (2014). Annex 2 Country Profiles. Available from: http://www.who.int/tb/data. [Last accessed on 2015 Jul 16].
Nyamogoba HD, Mbuthia G, Mining S, Kikuvi G, Biegon R, Mpoke S, et al. HIV co-infection with tuberculous and non-tuberculous mycobacteria in Western Kenya: Challenges in the diagnosis and management. Afr Health Sci 2012;12:305-11.
Siddiqi SH, Gerdes SR. MGITTM Procedure Manual. Mycobacteria Growth Indicator Tube (MGIT). Culture and Drug Susceptibility Demonstration; 2006. p. 15-6.
Manual for Sputum Smear Microscipy. Revised National Tuberculosis Control Programme. Central TB Division, Directorate General of Health Services, Ministry of Health and Family Welfare, Nirman Bhavan, New Delhi. Available from: AQ2 ???. [Last accessed on 2015 Aug 08].
Revised National TB Control Programme: Manual of Standard Operating Procedures (SOPs). Culture of Mycobacterium tuberculosis and drug Susceptibility Testing on Solid Medium. Version No. 01.01. New Delhi: Central TB Division, Directorate General of Health Services, Ministry of Health and Family Welfare, Nirman Bhavan; 2009.
Kim BJ, Park JH, Lee SA, Kim H, Cha CY, Kook YH, et al. Differentiation of mycobacteria in sputa by duplex polymerase chain reaction for mycobacterial hsp65 gene. Diagn Microbiol Infect Dis 2008;62:193-8.
Nath SS. Serological immune diagnosis of tuberculosis using PCR and in-house developed clinical isolate based ELISA. Int J Pharm Pharm Sci 2012;5(3):584-92.
Wang RF, Shi C, Lu M, Fang Z, Lu J, Wang F, et al. An improved whole-blood gamma interferon assay based the CFP21-MPT64 fusion protein. Clin Vaccine Immunol 2009;16:686-91.
Kumar VG, Urs TA, Ranganath RR. MPT 64 Antigen detection for rapid confirmation of M. tuberculosis isolates. BMC Res Notes 2011;4:79.
Shenoy VP, Mukhopadhyay C. Rapid immunochromatographic test for the identification and discrimination of Mycobacterium tuberculosis complex isolates from non-tuberculous mycobacteria. J Clin Diagn Res 2014;8(4):DC13-5.
Marzouk M, Kahla IB, Hannachi N, Ferjeni A, Salma WB, Ghezal S, et al. Evaluation of an immunochromatographic assay for rapid identification of Mycobacterium tuberculosis complex in clinical isolates. Diagn Microbiol Infect Dis 2011;69(4):396-9.
Martin A, Bombeeck D, Mulders W, Fissette K, De Rijk P, Palomino JC. Evaluation of the TB Ag MPT64 rapid test for the identification of Mycobacterium tuberculosis complex. Int J Tuberc Lung Dis 2011;15(5):703-5.
Maurya AK, Nag VL, Kant S, Kushwaha RA, Kumar M, Mishra V, et al. Evaluation of an immunochromatographic test for discrimination between Mycobacterium tuberculosis complex and non tuberculous mycobacteria in clinical isolates from extra-pulmonary tuberculosis. Indian J Med Res 2012;135(6):901-6.
SD Bioline TB Ag MPT64 - Is it TB or Non TB. Available from: Http:// www.alere.es/UserFiles/TBAgMPT64.pdf.
Hillemann D, Rüsch-Gerdes S, Richter E. Application of the Capilia TB assay for culture confirmation of Mycobacterium tuberculosis complex isolates. Int J Tuberc Lung Dis 2005;9(12):1409-11.
Saranya P, Swathi S, Kousalya K, Praveen D. A prospective interventional study of knowledge, attitude and practice (KAP) towards tuberculosis among patients with Koch’s disease. Int J Pharm Pharm Sci 2016;8:58-61.
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