DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20162919

Evaluation of various culture and staining techniques for the detection of extra pulmonary tuberculosis

Seema Gupta, Rahul K. Goyal, Rajesh Bareja, Rabindra N. Behara

Abstract


Background: Though pulmonary tuberculosis form is the commonest presentation, the extra pulmonary tuberculosis (EPTB) is also an important emerging clinical problem. The objective of the current study was to compare two staining techniques, Ziehl-Neelsen (ZN) stain, fluorescent stain and two-culture medium, solid Löwenstein-Jensen (LJ) medium and liquid 7H9 Middle brook medium in MGIT (Mycobacterium Growth Indicator Tube) 320 system, for detection of Mycobacterium in clinically suspected patients of EPTB.

Methods: A total of 100 clinically suspected cases of EPTB samples from various extrapulmonary sites had been collected. All the specimens were stained with ZN stain and fluorescent stain. The culture were processed after decontamination of specimens with NaOH-NALC method and thereafter inoculated on solid and liquid culture medium.

Results: Out of the 100 EPTB specimens, 30 were found positive by any of the above techniques used. Out of 30 positive cases 18 showed positivity by ZN staining while 20 showed positivity by fluorescent staining technique. In two culture methods, 27 isolates were grown by any of the culture system. Out of 27, 22 and 26 specimens showed growth of MTB complex on LJ media and MGIT culture system respectively. In AFB smear positive specimens, the average turnaround time was found to be 8.45 days and 22.5 days in MGIT and LJ medium culture assay respectively. While the turnaround times in AFB smear negative cases, it was 16.5 days and 32.3 days in MGIT and LJ medium culture assay respectively.

Conclusions: MGIT was a dependable, highly efficient system for recovery of MTB complex for EPTB specimens in combination with LJ media.


Keywords


EPTB, Fluorescent staining, LJ media, MGIT 320, ZN staining

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References


WHO global tuberculosis report 2015. www.who.int/tb/publications/global_report/en/.

Sharma SK, Mohan A. Extrapulmmonary tuberculosis-Review article. Indian J Med Res. 2004;120:316-53.

Negi SS, Khan SF, Gupta S, Pasha ST, Khare S, Lal S. Comparison of conventional diagnostic modalities, BACTEC culture and PCR test for diagnosis of tuberculosis. Indian J Med Microbiol. 2005;23:29-33.

Pottathil S, Nair A, Singh V, Kumar S, Bareja R. Evaluation of rapid techniques for the detection of mycobacteria in sputum with scanty bacilli or clinically evident, smear negative cases of pulmonary and extra-pulmonary tuberculosis. Mem Inst Oswaldo Cruz. 2011; 106 (5): 620-24.

Kent PT, Kubica GP. 1985. Public health mycobacteriology - a guide for the level III laboratory, Department of Health and Human Services/Centers for Disease Control and Prevention, Atlanta. 207.

Githui W, Kitui F, Juma ES, Obwana DO, Mwai J, Kwamasga D. A comparative study on reliability of the fluorescent microscopy and Ziehl Neelsen method in the diagnosis of pulmonary tuberculosis. East Afr Med J. 1993;70:263-6.

Ulukanligil M, Aslan G, Tasci S. A comparative study on the different staining methods and number of specimens for the detection of acid fast bacilli. Mem Inst. Oswaldo Cruz. 2000;95:855-8.

Murray SJ, Barrett A, Magee JG, Freeman R. Optimisation of acid fast smears for the direct detection of Mycobacteria in clinical samples. J Clin Path. 2003;56:613-5.

Prasthani K, Kumari AR. Efficacy of fluorochrome stain in the diagnosis of pulmonary tuberculosis coinfected with HIV. Ind J Med Microbiol. 2005;23(3):179-85.

Jain A, Bhargawa A, Agarawal SK. A comparative study of two commonly used staining techniques for acid fast bacilli in clinical specimens. Ind J Tub. 2002;49:161.

Rishi S, Sinha P, Malhotra B. A comparative study for the detection of Mycobacterium by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination. Ind J Med Microbiol. 2007;25(4);383-6.

Lu D, Heeren B, Dunne WM. Comparison of the automated MGIT with Lowenstein Jensen media for recovery of Mycobacteria from clinical specimens. Am J Clin Pathol. 2002;118:542-45.

Hanna BA, Ebrahimzadeh A, Elliot LB, Morgan MA, Novak SM, Rusch-Gerdes S, et al. Multicentre evaluation of BACTEC MGIT960 system for recovery of Mycobacteria. J Clin Microbiol. 1999;37:748-52.

Alcaide F, Benitez MA, Escriba JM, Martin R. Evaluation of BACTEC MGIT 960 and the MB/BacT systems for recovery of Mycobacteria from clinical specimens and for species identification by DNA Accu Probe. J Clin Microbiol 2000;38:398-01.

Tortoli E, Cichero P, Pietsimoni C, Simonetti MT, Gesu G, Nista D. Use of BACTEC MGIT 960 for recovery of Mycobacteria from clinical specimens:Multicentre study. J Clin Microbiol. 1999;17:578-82.

Idiogras P, Beristain X, Iturzaeta A, Vicente D, Perez Trallero E. Comparison of the automated non radiometric BACTEC MGIT 960 system with Lowenstein Jensen media. Colestos, MB 7H11 solid media for recovery of Mycobacteria. Eur J Clin Microbiol Infect Dis. 2000;19:350-54.

Somoskovi A, Kodman C, Lantos A, Bartfai Z, Tamasi L, Fuzi J, et al. Comparison of recoveries of Mycobacterium tuberculosis using the automated BACTEC MGIT960 system, BACTEC 460 TB system and Lowenstein Jensen media. J Clin Microbiol. 2000;38:2395-397.

Huang TS, Chen CS, Lee SS, Huang WK, Liu YC. Comparison of BACTEC MGIT960 and BACTEC460 TB system for the detection of Mycobacteria in clinical specimens. Am Clin Lab Sci. 2001;31:279-83.