Research Article
Reprocessing of Contaminated MGIT 960 Cultures to Improve
Availability of Valid Results for Mycobacteria
Balaji Subramanyam, Gomathi Sivaramakrishnan, Devi Sangamithrai, Rajkumar Ravi,
Kannan Thiruvengadam, Vaishnavi Vijayaragavan, Ayswarya Selvaraju,
Srikanth Prasad Tripathy, and Rajesh Mondal
Department of Bacteriology, ICMR-National Institute for Research in Tuberculosis, Chennai, India
Correspondence should be addressed to Rajesh Mondal; rajesh.m@nirt.res.in
Received 20 November 2019; Revised 18 June 2020; Accepted 30 June 2020; Published 18 July 2020
AcademicEditor:ToddR.Callaway
Copyright © 2020 Balaji Subramanyam et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Optimal recovery of mycobacteria from the contaminated liquid culture is a challenge. While alternative methods have been
suggested to reduce the rate of contamination in the BACTEC MGIT 960 system, reprocessing the contaminated liquid culture
improves recovery of Mycobacterium tuberculosis. Among 793 MGIT cultures raised from as many sputum specimens after
primarydecontaminationbythestandardNaLC-NaOHmethod,validresultswereavailablefor687(86.6%)as106(13.4%)were
contaminated.Reprocessingandreculturingofthecontaminatedculturesincreasedvalidresultsto739(93.2%)andreducedthe
contamination rate to 6.8%. Both values were statistically significant. Recovery of the Mycobacterium tuberculosis complex
increasedfrom45.6%to48.4%.Validnegativeresultswereavailableforanadditional3.4%.emethodmaybeadoptedtoreduce
the rate of contamination and to improve the valid culture results for mycobacteria.
1. Introduction
e Center for Disease Control and Prevention (CDC)
recommends the use of most efficient and rapid methods
availablefordiagnosisoftuberculosis[1].eyincludethe
use of at least one liquid medium such as the BACTEC
MGIT 960 method for the primary isolation of myco-
bacteria [2]. Controlling the overgrowth of normal flora
present in sputum is a major challenge while culturing
mycobacteriaintheliquidmedium.Overgrowthofnormal
floraleadstocontaminationwhichaffectsthesensitivityof
detection of the method especially in temperate settings.
BACTEC MGIT 960 is currently considered as the gold
standardbytheWorldHealthOrganization(WHO)andis
used widely for primary isolation as well as drug suscep-
tibility testing (DST) of Mycobacterium tuberculosis
(MTB).However,highratesofcontaminationinMGIT960
[3] are known to result in lower recovery rate of myco-
bacteria [4].
Alternatives such as use of a higher concentration of
NaOH up to 1.5% for decontamination of sputum and
increasing the volume of the antimicrobial supplement
PANTA have been suggested by the manufacturer for re-
ducing the rate of contamination. Reprocessing of the
contaminated culture and incubating the same are also
recommended but rarely practiced [5]. Failure to recover
MTB hampers DST and raises concern especially if the
cultureiscrucialforpatientmanagementandalsointhecase
of extrapulmonary specimens.
e present work aims to report on reprocessing of the
contaminated liquid culture for optimal recovery of
mycobacteria and to reduce the rate of contamination from
clinical specimens of a temperate endemic setting.
2. Materials and Methods
e observational study was done at the National Institute
for Research in Tuberculosis (NIRT), Chennai, India, and
was based on retrospective analysis of liquid cultures of all
diagnostic and follow-up sputum specimens collected from
patients under the NIRT Institutional Ethics Committee-
Hindawi
International Journal of Microbiology
Volume 2020, Article ID 1721020, 3 pages
https://doi.org/10.1155/2020/1721020