This is the very first step in Culture specimen for diagnosis and identification Tuberculosis. Sputum decontamination is the process of removing contamination, particularly the cleaning off of dangerous materials (sputum). Sputum specimen was collected in falcon 50 ml in 2 collection times ie morning an in place/health provider (labeled with P and S). In this study, we examine sputum smear negatives sent by 3 BP4 (Balai Pengobatan Penyakit Paru-Paru) in Yogyakarta.
The decontamination is processed in the Basic Safety Containment level 2 (BSC 2) to protect the laboratory workers from the infection risks, since the transmission of TB can be through aerosol. The lab workers should wear the protection equipments: lab jacket, gloves and mask. The steps are described below:
1. Mix NaOH 3% and Na Citrat with the same volume (25+25 ml)
2. Add NALC 0.25 gr (after mix with NaOH-Na Citrat should be used within 24 hours)
this is known as decontamination solution.
3. Add the solution into specimen with volume 1:1
4. mix in vortex mixer
5. Add PBS (Phosphate Buffer Saline) until volume is 45 ml
6. centrifuge in 4 degree, 3500 RPM for 15 minutes.
7. Throw the all of the supernatant and add with PBS 2ml
8. Mix with vortex mixer
The sputum has been decontaminated and centrifuged and ready to be inoculated in the medium. In this study the decontaminated specimen is inoculated in Thin Layer Agar (with and without Para Nitro Benzene/PNB), Lowenstein Jensen (with and without PNB) Mycobacteria Growth Indicator Tube (MGIT) and object glass for direct microscopic examination. The rest of the specimen is kept in the pellet and freeze in -20 degree to be used for other purposes examination.
The decontamination is processed in the Basic Safety Containment level 2 (BSC 2) to protect the laboratory workers from the infection risks, since the transmission of TB can be through aerosol. The lab workers should wear the protection equipments: lab jacket, gloves and mask. The steps are described below:
1. Mix NaOH 3% and Na Citrat with the same volume (25+25 ml)
2. Add NALC 0.25 gr (after mix with NaOH-Na Citrat should be used within 24 hours)
this is known as decontamination solution.
3. Add the solution into specimen with volume 1:1
4. mix in vortex mixer
5. Add PBS (Phosphate Buffer Saline) until volume is 45 ml
6. centrifuge in 4 degree, 3500 RPM for 15 minutes.
7. Throw the all of the supernatant and add with PBS 2ml
8. Mix with vortex mixer
The sputum has been decontaminated and centrifuged and ready to be inoculated in the medium. In this study the decontaminated specimen is inoculated in Thin Layer Agar (with and without Para Nitro Benzene/PNB), Lowenstein Jensen (with and without PNB) Mycobacteria Growth Indicator Tube (MGIT) and object glass for direct microscopic examination. The rest of the specimen is kept in the pellet and freeze in -20 degree to be used for other purposes examination.
