Selective Supplement for Burkholderia cepacia (BCSA)

Description

The detection of Burkholderia cepacia is important in water systems, particularly where the water is to be used for the preparation of pharmaceuticals and cosmetics. The organism is resistant to many commonly-used disinfectants.

Technique

Collect, dilute and prepare samples and volumes as required according to specifications, directives, official standard regulations and/or expected results.

Once solidified on a flat surface, spread the plate by streaking methodology or by spiral method.

Reconstitute the vial with 6 mL of sterile distilled water in aseptic conditions and add it to 500 mL of sterilized BCSA Agar Base cooled to 50 °C.

Do not overheat once supplemented.

Once solidified on a flat surface, spread the plates by streaking methodology or by spiral method.

Incubate the plates right side up aerobically at 33-37 °C for 48-72 h.

(Incubation times longer than those mentioned above or different incubation temperatures may be required depending on the sample, on the specifications, etc.)

After incubation, enumerate all the colonies that have appeared on the surface of the agar.

Each laboratory must evaluate the results according to their specifications.

Precautions

Do not reuse. For professional use only.

Do not use the product if it shows evidence of microbial contamination, discoloration, drying, cracking or other signs of deterioration.

Reconstitute the original freeze-dried vial by adding 6 mL of sterile distilled water.

Note: each vial is sufficient to supplement 500 mL of medium base for Burkholderia cepacia spp.

Physical/chemical control

Microbiological control

Reconstitute 1 vial as indicated in Composition; shake and dissolve completely.

Add 1 vial to 500 mL of medium base. DO NOT HEAT once supplemented.

Analytical methodology according to ISO 11133:2014/Amd 1:2018; A2:2020.

Aerobiosis. Incubation at 30-35 °C. Reading at 24-48 h, up to 72 h.

According to ISO 11133 and USP Pharmacopeia.

Sterility control

Add 5 mL of the sample to 100 mL of TSB and to 100 mL of Thioglycollate.

Incubation 48 hours at 30-35 °C and 48 hours at 20-25 °C: NO GROWTH.

Check at 7 days after incubation in same conditions.

• COENYE, T., P. VANDAMME, J.R.W. Govan and J.J. LiPuma. 2001. Taxonomy and identification of the Burkholderia cepacia Complex. J. Clin. Microbiol. 39:10:3427-3436.
• ERAM, S.M., Q.B. NEJAD, G.R. Khatami, N. NAFISSI. 2004. Detection of Burkholderia cepacia Complex in Patients with cystic fibrosis. Tanaffos 3:9:47-52.
• GILLIGAN, P.H., P.A. GAGE, L.M. BRADSHAW, D.V. SCHIDLOW and LV DeCicco. 1985. Isolation medium for the recovery of Pseudomonas cepacia from respiratory Secretions of Patients with cystic fibrosis. J. Clin. Microbiol. 22:5-8.
• HENRY, d.Ä., M.E. CAMPBELL, J.J. LiPuma and D.P. SPEERT. 1997. Identification of Burkholderia cepacia Isolates from Patients with cystic fibrosis and use of a simple new selective medium. J. Clin. Microbiol. 35:3:614-619.
• HENRY, d.Ä., M.E. CAMPBELL, C. McGimpsey, A. CLARKE, L. Louden, J.L. BURNS, M.H. ROE, P. VANDAMME and D.P. SPEERT. 1999. Comparison of isolation media for recovery from Burkholderia cepacia Complex Respiratory Secretions of Patients with cystic fibrosis. J. Clin. Microbiol. 37:4:1004-1007.
• ISLES, A., I. MACLUSKY, M. COREY, R. GOLD, C. Prober, P. FLEMING, and H. LEVISON. 1984. Pseudomonas cepacia in cystic fibrosis Infection: An Emerging problem. J. Pediatr. 104:206-210.
• MILLER, JJ, 1996. A guide to Specimen Management in Clinical Microbiology. ASM Press., Washington D.C.
• WELCH, DF, M.J. Muszynski, C.H. PAI, M.J. MARCON, M.M. Hribar, P.H. Gilligan, J.M. Matsen, P.G. Ahlin, B.C. HILMAN and SA CHARTRAND. 1987. Selective and differential medium for recovery from the Pseudomonas cepacia respiratory tracts of Patients with cystic fibrosis. J. Clin. Microbiol. 25:1730-1734.
• ISO Standard 11133:2014 Microbiology of food, animal feed and water. Preparation, production, storage, and performance testing of culture media.
• USP 42 - NF 37 1S (2019) Chapter <60> Microbiological examination of nonsterile products: Test for Burkholderia cepacia complex. USP Corp. Inc. Rockville. MD. USA.