DOC # CS-100656 REV 0

Product Insert TRYPTIC SOY BLOOD AGAR (TSBA) Products: AS-542

Tryptic Soy Blood Agar (TSBA)

4 plates / pkg

Intended Use TSBA is intended for the isolation and cultivation of many fastidious and non-fastidious microorganisms found in clinical specimens. This medium is also suitable for use in the CAMP test for the presumptive identification of group B Streptococci.

Summary TSBA is an enriched non-selective medium, supplemented with sheep blood for the purpose of improving - hemolytic reactions of Clostridium perfringens. It is supplemented with hemin and vitamin K1 to improve the recovery of fastidious anaerobes. This medium is prepared, stored and dispensed under oxygen-free conditions to prevent the formation of oxidized products prior to use.

Formula (g/L) Pancreatic Digest of Casein Soy Peptone Sodium Chloride Hemin (0.1% solution) Vitamin K1 (1.0% solution) Agar Sheep Blood DI Water

15.0 g 5.0 g 5.0 g 5.0 mL 1.0 mL 15.0 g 45.5 mL 1.0 L

Final pH 7.1 +/- 0.2 at 25 degrees C. Final weight 16.0 g +/- 1.6.

Precautions For IN VITRO DIAGNOSTIC USE only. Approved biohazard precautions and aseptic techniques should be observed when using this product. This product is for use only by properly-trained and qualified personnel. Sterilize all biohazard waste prior to disposal.

Storage and Shelf Life Storage: Upon receipt, store at room temperature (13°C - 27°C) in original container until use. Avoid overheating or freezing. Do not use medium if there are signs of deterioration (shrinking, cracking or discoloration due to oxidation of media) or contamination. The expiration date applies to the product in its original packaging and stored as directed. Do not use product past the expiration date shown on the container. Shelf Life: 90 days from date of manufacture.

Procedure Specimen Collection: Specimens for anaerobic culture should be protected from oxygen during collection, transportation and processing. Consult appropriate references for detailed instructions concerning collection and transportation of anaerobes. Methods for Use: TSBA should be inoculated directly with clinical material or a broth that has been previously inoculated from clinical material. Inoculated plates should be streaked to obtain isolated colonies, immediately placed in an anaerobic atmosphere and incubated at 35-37oC for 18-48 hours. Additional periods of incubation may be necessary to recover some anaerobes. Detailed instructions for processing anaerobic cultures can be found in the appropriate references.

Materials Required But Not Provided Standard microbiological supplies and equipment such as loops, saline blanks, slides, staining supplies, microscope, incinerator / autoclave, incubators, anaerobic chamber or anaerobic jars, disinfectant, other culture media and serological and biochemical reagents.

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Anaerobe Systems 15906 Concord Circle Morgan Hill, CA 95037 http://www.anaerobesystems.com

408 782 7557 Fax 408 782 3031

DOC # CS-100656 REV 0

Limitations TSBA will not provide complete information for identification of bacterial isolates. Additional test procedures and media are required for complete identification. This medium may not support the growth of all clinically significant anaerobes. Consult reference materials for additional information.

Quality Control TSBA should support good growth of fastidious microorganisms found in clinical infections. In addition, this media should support typical double zone of ß- hemolysis around colonies of Clostridium perfringens. The following organisms are routinely used for quality control testing at Anaerobe Systems. Organism Tested ATCC # Results Time Special Reaction (Hours) Bacteroides fragilis ** 25285 Growth 24 Prevotella melaninogenica ** 25845 Growth 48 Fusobacterium nucleatum ** 25586 Growth 24 – 48 Clostridium perfringens ** 13124 Growth 24 Double Zone of ß-hemolysis Peptostreptococcus anaerobius ** 27337 Growth 24 – 48 Staphylococcus aureus or 25923 Growth 24 Enterococcus faecalis 29212 Escherichia coli 25922 Growth 24 Proteus mirabilis 12453 Growth 24 Propionibacterium acnes or 6919 Growth 24 – 48 Clostridium difficile 9686 24 – 48 ** Organisms specified by NCCLS for quality control of Anaerobic Blood Agars User Quality Control: The final determination to the extent and quantity of user laboratory quality control must be determined by the end user. If sterility testing is to be performed on this product, the appropriate percentage of the original shipment amount should be incubated anaerobically and aerobically for 48 – 96 hours. If the nutritive capacity of this medium is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth. Organism B. fragilis C. perfringens P. anaerobius E. faecalis

Growth in Hours 24 hrs 24 hrs 24 hrs 24 hrs

Expected Results Double zone of  -hemolysis -

Physical Appearance: TSBA should appear opaque burgundy red in color.

References 1.

Dowell, V. R., Jr. and T. M. Hawkins. 1974. Laboratory Methods in Anaerobic Bacteriology. CDC Laboratory Manual. USDHEW C. D. C. Atlanta, GA 30333.

2.

Holdeman, L. V., F. P. Cato and W. E. C. Moore. 1977. Anaerobe Laboratory Manual. Virginia Polytechnic Institute and State University. Blacksburg, VA 24061.

3.

Somer- Jousimies, H. R., Summanen, P., Citron, D. M., Baron, E. J., Wexler, H. M. and S. M. Finegold. 2002. Wadsworth – KTL Anaerobic Bacteriology Manual. Star Publishing Co., Belmont, CA 94002.

4.

NCCLS. Quality Control for Commercially Prepared Microbiological Culture Media; Approved Standard- Third Edition. (2004). NCCLS document M22-A3. NCCLS, 940 West Valley Road, Suite 1400, Wayne, PA 19087-1898.

5.

Engelkirk, P. G., Duben-Engelkirk, J. and Dowell, V. R. 1992. Principles and Practices of Clinical Anaerobic Bacteriology. Star Publishing Co., Belmont, CA 94002.

Revision Date: 9/1/15

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Anaerobe Systems 15906 Concord Circle Morgan Hill, CA 95037 http://www.anaerobesystems.com

408 782 7557 Fax 408 782 3031