MGIT™ 960 PZA Preparation & Inoculation

From a Positive 7 mL MGIT Tube:

A. Day of Positivity = Day 0

Incubate at least one more day after
A
positivity.
Day 0
Incubate for one day C B. If tube is ‘Day 1or Day 2’, go directly to
Preparation & Inoculation.
Day 1 or 2 B Days 3-5
No Dilution required 1:5 dilution C. If tube is ‘Day 3’ to ‘Day 5’, dilute 1 mL of
positive broth into 4 mL sterile saline. Mix.

PZA Inoculum

From Solid Media: 1) Use growth on solid medium < 14 days old.
2) Make suspension >1.0 McFarland turbidity
standard in MGIT broth containing 8 - 10 glass
beads.
3) Vortex 2 - 3 minutes.
4) Allow to sit 20 minutes for large clumps to settle.
5) Transfer supernatent fluid to another sterile tube
Sterile Tube with and allow to sit for another 15 minutes.
4 mL MGIT broth
Solid Medium with and glass beads
6) Transfer supernatent fluid to another sterile tube
growth < 14 days old and adjust turbidity to 0.5 McFarland turbidity
standard.
7) Dilute 1 mL of final suspension in 4 mL sterile
Prepare suspension
> 1.0 McFarland
saline. Mix.
Vortex 2 - 3 minutes

Transfer Transfer
supernatent supernatent
1 mL

20 minutes for large 15 minutes for Adjust to 0.5 4 mL Sterile Saline

clumps to settle clumps to settle McFarland turbidity 1:5 dilution

PZA Inoculum

MGIT PZA Chart.ppt

Revision: New
October, 2000
 MGIT™ 960 PZA Preparation & Inoculation

0.8 mL
PZA Supplement
1. Add 0.8 mL BACTEC™ MGIT™ 960 PZA
Supplement to each tube.

Growth
PZA
Control

2. Add 100 µL of PZA drug to the tube labeled

100 µl Drug PZA only.

Growth
PZA
Control

3. Prepare Growth Control Inoculum: Pipette

0.5 mL of organism suspension into 4.5 mL
0.5 mL organism sterile saline to make a 1:10 dilution. Mix
suspension

4.5 mL Sterile Saline

4. Inoculate the Growth Control tube with 0.5

mL of 1:10 dilution. Mix

0.5 mL of 1:10 5. Inoculate the PZA drug containing tube with

dilution 0.5 mL of organism suspension (undiluted).
Mix

0.5 mL of 6. Place tubes into appropriate AST Set

Organism Carrier and enter into BACTEC MGIT 960
Suspension instrument.

7. Select PZA as the carrier set definition.

Growth
PZA
Control

MGIT PZA Chart.ppt

Revision: New
October, 2000