Bacterial Isolation of Mycobacterium avium
Tetsuya Komatsu, Kenji Ohya, Justice Opare Odoi, Shota Suganuma, Kotaro Sawai
Abstract
Mycobacterium avium subsp. hominissuis (MAH) is one of the most important agents causing non-tuberculosis mycobacterial infection in human and pigs. Genome analysis on MAH of human isolates has been proceeding, however, those of pigs are limited despite its potential source of infection to human. Here, we isolated MAH from pig lymph nodes or livers and obtained their genomes.
Steps
Cut out 20 mg of the caseous necrotic area and its surroundings from lymph nodes or organs and put them in a screw tube.
Put some 5 mm stainless steel spheres and 400 µl of 2% NaOH in the screw tube.
Set the tube in a tabletop crusher and process it at 3,200 rpm for 30 seconds.
Add 400 µl of 0.5% N-Acetyl-L cysteine (NALC)-2% NaOH and allow to stand for 14 hours.
Inoculate 100 µl of the suspension into the 2% Ogawa PS medium and incubate at 37°C for 3 - 4 weeks.
