Primary isolation of Mycobacterium

tuberculosis
using
BACTEC MGIT 960
DR. NOYAL MARIYA JOSEPH
Assistant Professor,
Dept. of Microbiology, JIPMER
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BACKGROUND:
The MGIT (Mycobacteria Growth Indicator
Tube) consists of 7 ml of modified
Middlebrook 7H9 broth medium

Growth Supplement (Oleic acid, Albumin,
Dextrose and Catalase), is added to support
the growth of mycobacteria, especially those
belonging to M. tuberculosis complex.

Addition of the MGIT PANTA is necessary to
suppress contamination.
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OBJECTIVE
At the end of the demonstration, you will learn:
the procedure for primary isolation of
Mycobacterium tuberculosis using BACTEC
MGIT 960 system
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PRINCIPLE
Modified Middlebrook 7H9 liquid media in MGIT
contains oxygen-quenched fluorochrome embedded in
silicone [tris 4, 7-diphenyl-1, 10-phenonthroline
ruthenium chloride pentahydrate]

During bacterial replication, free oxygen is utilized.
Depletion of oxygen results in increasing fluorescence
within the MGIT tube

MGIT 960 is an automated system which maintains an
incubation temperature of 37C and monitors for
increasing fluorescence every 60 minutes
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Preparation of NALC NaOH Trisodium citrate
solution
Preparation of Phosphate Buffered Saline
Addition of 0.8 ml of PANTA OADC
supplements into MGIT 7 ml tubes
Sterile work
Digestion, Decontamination and
Concentration
Inoculation of sample into MGIT 7 ml tube, LJ
slants
Preparation of concentrated smear
Specimen
processing
PROCEDURE
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SPECIMEN
DIRECT SMEAR MGIT 960
NEGATIVE POSITIVE
BLOOD AGAR SUBCULTURE
GROWTH
CONTAMINATED
NO GROWTH
PURE CULTURE
ZN STAINING
POSITIVE FOR
AFB
ICT / PNB
M.tuberculosis MOTT
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PC
H37RV
TEST
TEST
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ADVANTAGES & DISADVANTAGES
Advantages
Automated
Non Radioactive
Non Invasive
High recovery rate
Less TTD
High capacity
Real-time monitoring
DST result in days than
weeks
storage of patient
details is possible
HIS /LIS

Disadvantages
Expertise is required
Moderate cost when
compared with the low
cost conventional LJ and
with the high end PCR
technique.
High contamination
rates.
Power and cooling is
necessary round the
clock.

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THANK YOU
10/9/2014 XIII Workshop