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The qualitative BACTEC™ 12B Mycobacteria Medium is recommended for the culture and recovery of mycobacteria from clinical specimens, sputum, gastric, urine, tissue, mucopurulent specimens, other body fluids and other respiratory secretions, differentiation of the Mycobacterium tuberculosis complex from other mycobacteria and drug susceptibility testing of M. tuberculosis.

The BACTEC Radiometric Technique has been widely used for the rapid recovery of mycobacteria from sputum and other clinical specimens.^1-4 All types of clinical specimens, pulmonary as well as extra-pulmonary, can be processed for BACTEC primary isolation in a manner similar to the conventional procedures.^2,5-7 Details are provided in the CDC Manual, Procedure for the Isolation and Identification of Mycobacteria.³

The sample to be tested is inoculated into one or more 12B Mycobacteria Medium vials, with a syringe through the rubber septum, and incubated. The culture vial is periodically placed into the BACTEC 460TB System instrument for testing, which consists of aspiration of the head space gas and assay of its radioactive content. A positive reading indicates the presence of viable microorganisms in the vial.

DO NOT USE vials past their expiration date.

DO NOT USE vials that exhibit any cracks or defects; discard the vial in an appropriate manner.

Quality Control Certificates are provided with each carton of media. Quality Control Certificates show test organisms, including ATCC™ cultures specified in the NCCLS standard, Quality Assurance for Commercially Prepared Microbiological Culture Media.¹⁸

As part of the daily maintenance schedule, Performance Test Vials (BACTEC Performance Test Kit) should be tested on the BACTEC instrument. If low readings are obtained, yet Performance Test Vials were tested properly, possible instrument problems may be indicated (see BACTEC PTK package insert PP-046JAA).

Quality control requirements must be performed in accordance with applicable local, state and/or federal regulations or accreditation requirements and your laboratory's standard Quality Control procedures. It is recommended that the user refer to pertinent NCCLS guidance and CLIA regulations for appropriate Quality Control practices.

For in vitro Diagnostic Use.

This product contains dry natural rubber.

Pathogenic microorganisms including hepatitis viruses and Human Immunodeficiency Virus, may be present in clinical specimens. "Standard Precautions"^8-11 and institutional guidelines should be followed in handling all items contaminated with blood and other body fluids.

Perform all procedures, including processing of specimens, smear preparation, inoculum preparation, making dilutions, inoculations of media, subculturing, etc., in a suitable biological safety cabinet in a room with an appropriate ventilation system as recommended by the CDC.³ Use a proper protective gown, mask, and gloves while handling specimens and cultures of potential pathogens. Follow the CDC and OSHA recommendations.

Prior to use, each vial should be examined for evidence of damage, deterioration or contamination such as cloudiness, bulging or depressed septum. DO NOT USE any vial showing evidence of contamination. A contaminated vial could contain positive pressure. Vial contamination may not be readily apparent.

Vials displaying turbidity, contamination, or discoloration (darkening) should not be used. Vials that exhibit evidence of damage should be discarded prior to inoculation. On rare occasions, the glass bottle may be cracked and the neck may break during removal of the flip-off cap or in handling. Also, on rare occasions a vial may not be sealed sufficiently. In both cases, the contents of the vials may leak or spill, especially if the vial is inverted. If the vial has been inoculated, treat the leak or spill with caution, as pathogenic organisms/agents may be present. Before discarding, sterilize all inoculated vials by autoclaving.

Positive culture vials for subculturing or staining, etc.: Before sampling it is necessary to release gas which may build up due to microbial metabolism. Inoculation and sampling should be performed in a biological cabinet, and appropriate protective clothing, including gloves and masks should be worn. See Procedure section for more information on subculturing.

To minimize the potential of leakage during inoculation of specimen into culture vials, use syringes with permanently attached needles or securely fastened Luer-Lok™ brand tips.

The rubber septum of a vial should be disinfected each time the vial is entered. Clean the top of each vial with disinfectant, followed by 70% alcohol. Use an appropriate disinfectant for cleaning the work area and swabbing the vial septum and other surfaces. The CDC states: "With so many disinfectants available, it is important to consult the product brochures to make certain that the disinfectant is bactericidal for mycobacteria."¹²

Working with a syringe and needle requires special attention and safety precautions. Use syringes with permanently attached needles or securely fastened Luer-Lok brand tips. Handle the syringe with appropriate caution. Do not put the cover of the needle back after use. Discard syringes in sharps containers and handle the disposal containers carefully. Refer to the NCCLS Approved Guideline M29, "Protection of Laboratory Workers from Occupationally Acquired Infections."⁸