An integrated approach to detection of Mycobacterium tuberculosis using BD MGIT 320 system and other diagnostic modalities.
Keywords:
Liquid culture, Multi drug resistant tuberculosis, extrapulmonary tuberculosis, Line Probe Assay
Abstract
Background: Application of automated liquid culture system like Becton, Dickinson and Company (BD) MGIT 320 can shorten the recovery time of Mycobacteria in clinical diagnostic laboratories as compared to Lowenstein Jensen (LJ) medium. The present study aimed to compare the results of BD MGIT 320 culture system with that of LJ medium and also to evaluate the usefulness of molecular methods as an adjunct.Methods: Specimen from clinically suspected cases of tuberculosis were processed as per standard guidelines for smear microscopy, solid culture in LJ media, liquid culture in BD MGIT 320 system and Line probe assay. Polymerase chain reaction was performed on extracted DNA from MGIT positive specimen to detect IS6110 and MPB64 gene targets.Result: Out of total 83 specimens, a statistically significant higher isolation rate (25.3%, n=21) was shown by MGIT 320 system in comparison to LJ medium (12.04%, n=10). Line Probe Assay was positive in 37.3% (n=31) and direct microscopy was positive in only 13.25% (n=11) specimen. The average duration of detection was found to be more rapid by MGIT320 than LJ culture (18 days vs 24.1 days). Among the 21 MGIT positive specimens 7 were found positive for both IS6110 and MPB64 gene while 7 specimens were found positive only for IS6110.Conclusion: The automated liquid culture systems like BD MGIT 320 have a higher isolation rate and are more rapid as compared to solid media. But adoption of Line probe assay combined with MGIT culture will definitely yield rapid and efficient detection of Mycobacteria. DOI: 10.21276/AABS.1736References
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15. Rodrigues C, Shenai S, Sadani M, Sukhadia N, Jani M, Ajbani K et al. Evaluation of the bactec MGIT 960 TB system for recovery and identification of Mycobacterium tuberculosis complex in a high volume tertiary care centre. Indian J Med Microbiol [serial online] 2009; 27:217-221. [cited 2017 Aug 28] http://www.ijmm.org/text.asp?2009/27/3/217/53203
16. Bunger R, Singh VA, Avneet, Mehta S, Pathania D. Evaluation of BACTEC Micro MGIT with Lowenstein Jensen media for detection of Mycobacteria in clinically suspected patients of extra pulmonary tuberculosis in a tertiary care hospital at Mullana (Ambala). J Med Microb Diagn. 2013,2:2. https://www.omicsonline.org/evaluation-of-bactec-micro-mgit-with-lowenstein-jensen-media-for-detection-of-mycobacteria-in-clinically-suspected-patients-of-extra-pulmonary-tuberculosis-2161-0703.1000123.php?aid=12037
17. Chihota VN, Grant AD, Fielding K, Ndibongo B, Zyl AV, Muirhead D et al. Liquid versus solid culture for tuberculosis: performance and cost in a resource-constrained setting. Int J Tuberc Lung Dis. 2010;14(8):1024-1031. https://www.ncbi.nlm.nih.gov/pubmed/20626948
18. Horner PJ, Moss FM: Tuberculosis in HIV infection. Int J STD AIDS 1991, 2(3):162-167. https://www.ncbi.nlm.nih.gov/pubmed/1863645.
19. Jatana S. K., Nair M. N., Lahiri K. K., Sarin N. P. Polymerase chain reaction in the diagnosis of tuberculosis. Indian Pediatr. 2000; 37, 375–382. http://indianpediatrics.net/april2000/april-375-382.htm
20. Narayanan S, Parandaman V, Narayanan P, Venkatesan P, Girish C, Mahadevan S, Rajajee S. Evaluation of PCR using TRC4 and IS6110 primers in detection of tuberculous meningitis. J Clin Microbiol. 2001; 39, 2006–2008. https://www.researchgate.net/publication/12010310
21. Radhakrishnan I, Manju Y K, Kumar R A & Mundayoor S. Implications of low frequency of IS6110 in fingerprinting field isolates of Mycobacterium tuberculosis from Kerala, India. J ClinMicrobiol. 2001 ; 39, 1683. pubmedcentralcanada.ca/pmcc/articles/PMC88004/pdf/jm001683.pdf
22. Bhigjee AI, Padayachee R, Paruk H, Hallwirth-Pillay KD, Marais S, Connoly C. Diagnosis of tuberculous meningitis: Clinical and laboratory parameters. Int J Infect Dis. 2007;11(4):348–354. http://www.ijidonline.com/article/S1201-9712(06)00190-1/fulltext
23. Rafi W, Venkataswamy MM, Ravi V, Chandramuki A. Rapid diagnosis of tuberculous meningitis: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB-64 and 65 kDa antigen. J Neurol Sci. 2007;252:163–168. http://www.jns-journal.com/article/S0022-510X(06)00529-6/fulltext
2. Duque A, Lin SY, Desmond E, Rienthong S, Rienthong D, Boonin C.Evaluation of the BD Bactec MGIT 320 for detection of Mycobacteria and drug susceptibility testing of Mycobacterium tuberculosis. J. Clin. Microbiol 2013;51:3403–3405. http://dx.doi.org/10.1128/JCM.01357-13
3. Hanna B A, Ebrahimzadeh A, Elliott L B, Morgan M A, Novac S M, Gerdes S R et al. Multicenter evaluation of the BACTEC MGIT 960 system for recovery of mycobacteria. J Clin Microbiol 1999; 37: 748–752. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC84542
4. Somoskovi A, Kodmon C, Lantos A, Bartfai Z, Tamasi L, Fuzy J et al. Comparison of recoveries of Mycobacterium tuberculosisusing the automated BACTEC MGIT 960 system, the BACTEC 460 TB system, and Lowenstein-Jensen medium [Internet]. J Clin Microbiol. 2000 Jun;38(6):2395-2397 [cited 2017 Sep 26] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC86818
5. Dowdy D, Lourenço M, Cavalcante S, Saraceni V, King B, Golub J et al. Impact and Cost-Effectiveness of Culture for Diagnosis of Tuberculosis in HIV-Infected Brazilian Adults. PLoS ONE [Internet]. 2008; 3(12):e4057 [cited 26 September 2017] https://doi.org/10.1371/journal.pone.0004057
6. Kent PT, Kubica GP. Public health mycobacteriology: a guide for the level III laboratory. Atlanta, GA: U.S. Dept. of Health and Human Services, Public Health Service, Centers for Disease Control; 1988.
7. Revised National TB Control Programme. Manual of standard operating procedures for culture of Mycobacterium tuberculosis and drug susceptibility testing on solid medium [Internet], Version No. 01.01, 2009. [cited 16 September 2017] http://tbcindia.nic.in/WriteReadData/l892s/6995271860Training manual M tuberculosis C DST.pdf
8. Siddiqi, S. H., & Rüsch-Gerdes, S. For BACTEC™ MGIT 960™ TB System [Internet] 2006. [cited 16 September 2017]
https://www.finddx.org/wp-content/uploads/2016/02/mgit_manual_nov2006.pdf
9. Hain Lifescience, GmbH, Nehren. Germany. Genotype MTBDRplus TM, version 2.0. Instruction manual [Internet][cited 2017Sep26] http://www.hainlifescience.com/pdf
10. Sharma K, Sinha S, Sharma A, Prasad K, Rana S, Sharma M, et al. Multiplex PCR for rapid diagnosis of gastrointestinal tuberculosis. [Internet] J Glob Infect Dis 2013;5(2):49-53 [cited 2017 Sep 27] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3703210
11. Nolte, F. S., B. Metchock. Mycobacterium. In P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H. Yolken, ed. by. Manual of clinical microbiology. 6th ed. ASM Press, Washington, D.C.; 1995. p. 400–437.
12. Tortoli E, Cichero P, Piersimoni C, Simonetti T, Gesu G, Nistta D. Use of BACTEC MGIT for recovery of mycobacteria from clinical specimens: multicentric study. J Clin Microbiol. 1999;37: 3578- 3582. http://jcm.asm.org/content/37/11/3578.full.pdf
13. Dongsi L, Bobby H, Dunne W M. Comparison of the automated mycobacteria growth indicator tube system (BACTEC MGIT 960) with Lowenstein Jensen medium for recovery of mycobacteria from clinical specimens. Microbiol Infect Dis. 2002; 118:542-545. https://academic.oup.com/ajcp/article-pdf/118/4/542/4984895/ajcpath118-0542.pdf
14. Rishi S, Sinha P, Malhotra B, Pal N. A Comparative study for the detection of mycobacteria by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination. Ind J Med Microbiol. 2007 Oct; 25(4):383-386. https://www.ncbi.nlm.nih.gov/pubmed/18087090
15. Rodrigues C, Shenai S, Sadani M, Sukhadia N, Jani M, Ajbani K et al. Evaluation of the bactec MGIT 960 TB system for recovery and identification of Mycobacterium tuberculosis complex in a high volume tertiary care centre. Indian J Med Microbiol [serial online] 2009; 27:217-221. [cited 2017 Aug 28] http://www.ijmm.org/text.asp?2009/27/3/217/53203
16. Bunger R, Singh VA, Avneet, Mehta S, Pathania D. Evaluation of BACTEC Micro MGIT with Lowenstein Jensen media for detection of Mycobacteria in clinically suspected patients of extra pulmonary tuberculosis in a tertiary care hospital at Mullana (Ambala). J Med Microb Diagn. 2013,2:2. https://www.omicsonline.org/evaluation-of-bactec-micro-mgit-with-lowenstein-jensen-media-for-detection-of-mycobacteria-in-clinically-suspected-patients-of-extra-pulmonary-tuberculosis-2161-0703.1000123.php?aid=12037
17. Chihota VN, Grant AD, Fielding K, Ndibongo B, Zyl AV, Muirhead D et al. Liquid versus solid culture for tuberculosis: performance and cost in a resource-constrained setting. Int J Tuberc Lung Dis. 2010;14(8):1024-1031. https://www.ncbi.nlm.nih.gov/pubmed/20626948
18. Horner PJ, Moss FM: Tuberculosis in HIV infection. Int J STD AIDS 1991, 2(3):162-167. https://www.ncbi.nlm.nih.gov/pubmed/1863645.
19. Jatana S. K., Nair M. N., Lahiri K. K., Sarin N. P. Polymerase chain reaction in the diagnosis of tuberculosis. Indian Pediatr. 2000; 37, 375–382. http://indianpediatrics.net/april2000/april-375-382.htm
20. Narayanan S, Parandaman V, Narayanan P, Venkatesan P, Girish C, Mahadevan S, Rajajee S. Evaluation of PCR using TRC4 and IS6110 primers in detection of tuberculous meningitis. J Clin Microbiol. 2001; 39, 2006–2008. https://www.researchgate.net/publication/12010310
21. Radhakrishnan I, Manju Y K, Kumar R A & Mundayoor S. Implications of low frequency of IS6110 in fingerprinting field isolates of Mycobacterium tuberculosis from Kerala, India. J ClinMicrobiol. 2001 ; 39, 1683. pubmedcentralcanada.ca/pmcc/articles/PMC88004/pdf/jm001683.pdf
22. Bhigjee AI, Padayachee R, Paruk H, Hallwirth-Pillay KD, Marais S, Connoly C. Diagnosis of tuberculous meningitis: Clinical and laboratory parameters. Int J Infect Dis. 2007;11(4):348–354. http://www.ijidonline.com/article/S1201-9712(06)00190-1/fulltext
23. Rafi W, Venkataswamy MM, Ravi V, Chandramuki A. Rapid diagnosis of tuberculous meningitis: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB-64 and 65 kDa antigen. J Neurol Sci. 2007;252:163–168. http://www.jns-journal.com/article/S0022-510X(06)00529-6/fulltext
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2017-11-17
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