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BACTEC™ MYCO/F-Sputa culture media (modified Middlebrook 7H9 broth with CO₂) with the addition of BACTEC™ Supplement/F and BACTEC™ PANTA™/F antibiotic mixture, when appropriate, is used with the BACTEC™ Brand 9000MB flourescent series instrument as a qualitative procedure for the in vitro culture and recovery of mycobacteria. Acceptable specimens to be used are digested decontaminated clinical specimens and sterile body fluids other than blood.

Since the mid-1980s and spread of the AIDS epidemic, Mycobacterium tuberculosis (MTB) and Mycobacteria other than tuberculosis (MOTT), especially Mycobacterium avium complex (MAC), have become resurgent. From 1985 to 1992, the number of MTB cases reported increased 18%. Tuberculosis still kills an estimated 3 million persons a year worldwide, making it the leading infectious disease cause of death.¹ Between 1981 and 1987, AIDS case surveillances indicated that 5.5% of the patients with AIDS had disseminated nontuberculous mycobacterial infections, e.g., MAC. By 1990, the increased cases of disseminated nontuberculous mycobacterial infections had resulted in a cumulative incidence of 7.6%.² In addition to the resurgence of MTB, multidrug-resistant MTB (MDR-TB) has become an increasing concern. Laboratory delays in the growth, identification and reporting of these MDR-TB cases contributed at least in part to the spread of the disease.

The Centers for Disease Control and Prevention (CDC) have recommended that every effort must be made for laboratories to use the most rapid methods available for diagnostic mycobacteria testing. These recommendations include the use of both a liquid and a solid medium for mycobacterial culture.³

The BACTEC 9000MB System is designed for the rapid detection of mycobacteria in clinical specimens other than blood. The system includes a liquid culture medium (Myco/F-Sputa Culture Vial), a growth supplement (Supplement/F) and an antibiotic supplement (BACTEC PANTA/F). BACTEC Supplement/F contains growth enhancers for mycobacteria, and is also used to reconstitute BACTEC PANTA/F. BACTEC PANTA/F contains antimicrobial agents used to suppress the growth of contaminating or normal flora microorganisms which may survive the decontamination process and is recommended as an addition with all non-sterile specimens. Each vial contains a sensor which can detect decreases in the dissolved oxygen in the medium resulting from microorganism metabolism and growth. The sensor is monitored by the instrument for increasing fluorescence which is proportional to decreasing dissolved oxygen content. A positive reading indicates the presumptive presence of viable mycobacteria in the vial. Detection is limited to microorganisms that will grow in the medium.

Quality Control Certificates are provided with each carton of media.

It is recommended that each new shipment or lot of BACTEC MYCO/F-Sputa culture media be tested with the ATCC control organisms identified in the chart below as a positive control and an uninoculated vial as a negative control.

To prepare the positive control vial:

1. Grow the organism in 7H9 broth.

2. Prepare a #1 McFarland (= 10⁷ CFU/mL) suspension of the organism using sterile saline.

3. Dilute the suspension to 10⁴ CFU/mL using sterile saline.

4. Inoculate 1 mL of this suspension into a BACTEC MYCO/F-Sputa culture vial which has been supplemented with 2.0 mL of reconstituted BACTEC PANTA/F. Inoculum of 1 mL is to be used for Quality Control organisms only.

The positive and negative control vials should be scanned into the instrument and tested. The positive control vial should be detected as instrument positive within the range given in the above chart. The negative control should remain negative. If either of these vials do not give the expected results, do not use the media until you have contacted Technical Services at 1-800-638-8663 (United States only).

For information on quality control for the BACTEC 9000MB System, refer to the BACTEC 9000MB User's Manual (MA-0092).

The BACTEC MYCO/F-Sputa culture vials contain the following active ingredients prior to processing:

All BACTEC media are dispensed with added CO₂. Composition may have been adjusted to meet specific performance requirements.

Prior to inoculation of respiratory and non-sterile specimens, each 40 mL vial of BACTEC MYCO/F-Sputa medium requires the addition of 2.0 mL of BACTEC PANTA/F Antibiotic Supplement solution as reconstituted with BACTEC Supplement/F. Prior to inoculation of sterile body fluids other than blood, each 40 mL vial of BACTEC MYCO/F-Sputa medium must be supplemented with 2.0 mL of BACTEC Supplement/F. Please see the Package Inserts for BACTEC PANTA/F (PP-102) and BACTEC Supplement/F (PP-103) for additional information.

For in vitro diagnostic.

This Product Contains Dry Natural Rubber.

POTENTIALLY INFECTIOUS TEST SPECIMEN. Observe "Universal Precautions"^4,5 and institutional guidelines when handling and disposing of infectious materials.

Biosafety Level 2 practice, containment equipment, and facilities are recommended for preparing acid-fast stains and for culturing clinical specimens. For activities involving the propagation and manipulation of Mycobacterium tuberculosis or Mycobacterium species grown in culture, Biosafety Level 3 practice, containment equipment, and facilities are required as recommended by the CDC.⁶

Prior to use, each vial should be examined for evidence of contamination such as cloudiness, bulging or depressed septum, or leakage. On rare occasions, the glass bottle neck may be cracked and the neck may break during removal of the flip-off cap or in handling. Also, a vial may exhibit incomplete crimping of the cap, as indicated when the metal edge at the bottom of the cap is not uniformly rolled under the neck finish of the bottle. In both cases the contents of the vials may leak or spill, especially if the vial is inverted. DO NOT USE any vial showing evidence of contamination, damage or incomplete crimping.

To minimize the potential of leakage during inoculation of specimen into culture vials, use a tuberculin syringe with a 25-gauge permanently attached needle. A one-handed inoculation technique and a suitable vial holder should be employed to prevent accidental needle stick injury. Needles larger than 20-gauge must NOT be used. For supplement addition, pressure release or subculturing, use of larger needles may permanently damage the vial septum, and may cause leakage.

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

A contaminated vial could contain positive pressure. If a contaminated vial is sampled, gas and/or contaminated culture medium could escape from the vial, creating an aerosol hazard. Vial contamination may not be readily apparent.

Before discarding, sterilize all inoculated MYCO/F-Sputa vials by autoclaving.