DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20171925

Comparison of NALC-NAOH processing method with C18-carboxy propyl betaine method for the detection of mycobacterium in sputum

G. Amar Kumar, S. Chandrasekaran

Abstract


Background: Tuberculosis is an infectious disease which remains to be a major public health risk worldwide even after the availability of many highly sensitive diagnostic tools. Early case detection plays an important role in control of the disease which relies solely on the detection of acid-fast bacilli in clinical samples which is low sensitive. Prior decontamination of sputum sample may improve the detection of mycobacterium.

Methods: A prospective study was conducted with a total of (N=464) clinically suspected TB patient sputum samples which were collected and processed directly for AFB. From which AFB +ve samples were excluded. The direct AFB-ve (N=279) samples were processed using concentration (NALC, CB-18) and culture (LJ medium). After concentration, smears were examined under oil immersion for acid fast bacilli by ZN staining.

Results: Out of 464 sputum samples, 185 direct ZN stain +ve were excluded. Among the ZN-ve (N=279) samples, N= 48 (17.2%) were positive using NALC-NaOH method. The positivity has improved to N=69 (25%) using CB-18 and N=79 (28.3%) using culture (LJ medium).

Conclusions: CB-18 is sensitive than NALC-NaOH method and its sensitivity is almost comparable to the gold standard culture using LJ medium. Therefore we conclude that the identification of AFB using CB-18 concentration method can be used in resource limited health care setups which might help us to diagnose the TB at the earliest.


Keywords


Acid fast bacilli, C18-carboxypropylbetaine, Lowenstein-jensen, NALC-NaOH-N-acetyl-L-cysteine sodium hydroxide, Ziehl-neelsen staining

Full Text:

PDF

References


Uddin MK, Chowdhury MR, Ahmed S, Rahman MT, Khatun R, van Leth F, Banu S. Comparison of direct versus concentrated smear microscopy in detection of pulmonary tuberculosis. BMC research notes. 2013;6(1):291.

W.H.O Global tuberculosis report 2016. Available from: http://www.who.int/tb/ publications/ global_report/en/

Frieden T. Toman’s tuberculosis case detection, treatment, and monitoring. 2nd edition. world health organization geneva. 2004:334.

Stinson KW, Eisenach K, Kayes S, Matsumoto M, Siddiqi S, Nakashima S, et al. Mycobacteriology Laboratory Manual, global laboratory initiative advancing TB diagnosis. 1sted. April 2014.

Verma S, Dhole TN, Kumar M, Kashyap S. Novel Approach for Improving Sensitivity of Microscopic Detection of Acid-Fast Bacilli by Use of the ReaSLR Method. JCM. 2013;51(11):3597-601.

Laserson KF, Yen NT, Thornton CG, Mai VT, Jones W, An DQ et al. Improved sensitivity of sputum smear microscopy after processing specimens with C18-carboxypropylbetaine to detect acid-fast bacilli: a study of United States-bound immigrants from Vietnam. JCM. 2005;43(7):3460-2.

Scott CP, Filho DA, Queiroz FC, Thornton CG, Bishai WR, Fonseca LS, et al. Comparison of C18-carboxypropylbetaine and standard N-Acetyl-L-Cysteine-NaOH processing of respiratory specimens for increasing tuberculosis smear sensitivity in Brazil. JCM. 2002;40(9):3219-22.

Morcillo, Nora, Belén Imperiale, and Juan Carlos Palomino. New simple decontamination method improves microscopic detection and culture of mycobacteria in clinical practice. 2008;1:21-6.

Sharath BN, Shastri S. India's new TB diagnostic algorithm-far from reality?. Public Health Action. 2016;6(3):206.

Revised National TB Control Programme Training Manual for Mycobacterium Tuberculosis Culture and Drug susceptibility testing. Central TB Division Directorate General of Health Services. 2009.

Fine AE, O'Brien DJ, Winterstein SR, Kaneene JB. "An effort to isolate Mycobacterium bovis from environmental substrates during investigations of bovine tuberculosis transmission sites (cattle farms and wildlife areas) in Michigan, USA. 2011: 787181.

Thornton CG, Maclellan KM, Brink TL, Lockwood DE, Romagnoli M, Turner J, et al. Novel method for processing respiratory specimens for detection of mycobacteria by using C18-carboxypropylbetaine: blinded study. JCM. 1998;36(7):1996-2003.

Dundee L, Grant IR, Ball HJ, Rowe MT. Comparative evaluation of four decontamination protocols for the isolation of Mycobacterium avium subsp. paratuberculosis from milk. LAM. 2001;33(3):173-7.

Thornton CG, Cranfield MR, MacLellan KM, Brink Jr TL, Strandberg JD, Carlin EA et al. Processing Postmortem Specimens with-Carboxypropylbetaine and Analysis by PCR to Develop an Antemortem Test for Mycobacterium avium Infections in Ducks. JZWM. 1999:11-24.

Thornton CG, MacLellan KM, Stabel JR, Carothers C, Whitlock RH, Passen S. Application of the C18-carboxypropylbetaine specimen processing method to recovery of Mycobacterium avium subsp. paratuberculosis from ruminant tissue specimens. JCM. 2002;40(5):1783-90.

Centers for Disease Control and Prevention National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention Division of Tuberculosis Elimination Core Curriculum on Tuberculosis: What the Clinician Should Know. 6th ed. 2013.