BACTEC MGIT 960 CULTURE SYSTEM

FOR MYCOBACTERIA

David Lubasi
Biomedical Scientist

TUBERCULOSIS WORKSHOP
NDOLA
AUGUST 2004
BACKGROUND

 In Zambia, a four-fold increase in tuberculosis

notification rates was noted in the last decade and the
current notification rate stands at over 512 per 100,000
per year(WHO/MOH report 1999)

 Lusaka province has highest incidence in the

country,standing at 800/100,000/year (1996 MOH
report).
 NUMBER OF PATIENTS
YEAR
POSITIVES NEGATIVES TOTAL POSITIVITY

1998 2206 6577 8783 25.1%

1999 2847 12620 15467 18.4%

2000 1806 11059 12865 14.0%

2001 1321 7911 9232 14.3%

2002 1454 7929 9383 15.5%

 
2003 1794 10164 11958 15.0%
Statistics of patients received in TB lab (Source; TB laboratory annual reports)
 Diagnosis of tuberculosis has used a variety of clinical and
microbiological methods, which, until recently, have changed
very little since the discovery of the tubercle bacillus over a
century ago. (Reichman et al 1993).

 Sensitivity of direct Microscopy is improved by use of the

NaOcl method (Habeenzu et al 1998), but still not as sensitive
as culture.

 Conventional culture for Mycobacteria is slow (on average 4

weeks and maximum incubation of 9 weeks).
 Radiometric BACTEC 460 TB and non radiometric BACTEC
9000 MB are more efficient in recovery of Mycobacteria
(ZANETTI et al 1997), but these systems are too expensive for
most developing countries.

 Due to the heavy workload ,the UTH TB lab acquired the

BACTEC MGIT 960, 960 purchase of which was financed by
JICA through the HIV/AIDS and TB control project .
INTRODUCTION
 MGIT stands for Mycobacteria Growth Indicator
Tube, and 960 indicates the total number of culture
tubes it can hold at any given time

 The BACTEC MGIT 960 System is an in vitro

diagnostic instrument for rapid detection of
Mycobacteria in clinical specimens other than blood.

 The system is designed to meet the needs of medium

and high volume labs,capable of processing about 8,000
cultures per year.

 This system is simple, efficient , safe to use and

occupies small laboratory space.
 PRINCIPLE OF THE TEST / PROCEDURE
 The MGIT 7ml tube contains modified middle brook 7H9
broth .
 Culture tubes contain a fluorescent sensor at the bottom
which responds to the concentration of oxygen .
 Initial concentration of dissolved oxygen quenches the
emission from the compound, and little fluorescence can be
detected.
 Actively respiring micro organisms consume the oxygen
which allows the compound to fluorescence.
 METHODS

 Specimens are collected and transported like in ordinary

microscopy and culture for Mycobacteria to the TB
laboratory.
 Decontaminate and digested specimens with an equal volume
of 4% sodium hydroxide for 15 minutes vortexing every 5
minutes
 Transfer the Decontaminated sputum in to a 50ml corning
tube.
 Centrifuged at 3000rpm for 15 minutes and discard the
supernatant.
 Add phosphate-buffered saline (PBS pH 6.8) to the residue
raising the volume to 50 ml.
 Centrifuge at 3000 rpm for 15 minutes and discard the
supernatant.
 Add 0.5 ml of PBS pH6.8 to resuspend the pellets.
 Add 0.5 ml of resuspended deposit to the MGIT culture tube
 MGIT tubes are then incubated into the BACTEC MGIT 960
system until positivity is observed for positive samples or
upto 8 weeks for negative ones.
 Positive cultures are usually detected within 4 – 14 days
 When positive tubes are identified, they are removed and are
confirmed for Acid fastness (AFB), for species identification
and drug susceptibility testing and later storage of isolate.
NB
MGIT culture tubes contains 7ml of Middlebrook 7H9 and
0.8 ml of PANTA.
 PROTOCOL LENGTH OF MGIT Vs LJ
CONVENTIONAL METHOD (DAYS)

LJ METHOD MGIT

Average (+) 29.6 4.4

Normal range(+) 28-56 4 – 14

Negatives 56 – 63 42 – 56
 RESULTS OF MGIT Vs. LJ FROM OTHER
RESEARCHERS
CONVENTIONAL BACTEC
STUDY CULTURE   SYSTEM
  
Van Griethiuysen et al,1996 79.9% 95.9%

Zanetti et al, 1997 69.3 % 95.0 %

Jesus et al , 2001 46.6% 86.6%

Lu et al , 2002 45.5 % 71.1 %

* Recovery rates from positive samples

 OBSERVATIONS
 Most of the studies on evaluation of mycobacteria recovery
from the fluorometric BACTEC 960 and the radiometric
BACTEC 460 TB system have shown that they are more
sensitive in recovery of mycobacteria than the conventional
L-J and smear microscopy.

 Zannetti et al (1997) showed no significant difference between

the radiometric BACTEC 460 TB and the fluorometric
BACTEC 960 thus 91.9% positivity and 95.1% positivity
respectively. But use of radioisotopically labeled substrates
has inhibited universal application of the radiometric
BACTEC 460 TB system.
 This system is simple, efficient , safe to use and
occupies small laboratry space.
 In poor economies MGIT is relatively expensive to
acquire and sustain.
CONCLUSION.

 Diagnosis of TB depends on the isolation and

identification of Mycobacteria and the control
of MDR tuberculosis depends rapid sensitivity
results. Therefore use of sensitive methods and
faster culture methods like the BACTEC
MGIT 960 system is necessary.
THE END