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Tryptic Soy Blood Agar (TSBA)

Tryptic Soy Blood Agar (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. TSBA is an enriched non-selective medium, supplemented with sheep blood for the hemolytic reactions.  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.

     Anaerobe Systems PRAS Tryptic Soy Blood
                        Agar (TSBA) AS-542

Tryptic Soy Blood Agar Products

 Item #





TSBA mono plate 

4 plates 



Pancreatic Digest of Casein, 15.0 g

Soy Peptone, 5.0 g

Sodium Chloride, 5.0 g

Hemin (0.1% soln), 5.0 ml

Vitamin K1 (1% soln), 1.0 ml

Agar, 15.0 g

Sheep Blood, 45.5 ml

Distilled Water, 1000.0 ml


Final pH 7.1 +/- 0.2 at 25 degrees C.

Final weight 16.0 g +/- 1.6.



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.


Specimen Collection:  Specimens for anaerobic culture should be protected from air (oxygen) during collection, transport and processing.  Consult appropriate references for detailed instructions concerning collection and transport 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.


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 hemolysis around colonies of Clostridium perfringens.

The following organisms are routinely used for quality control testing at Anaerobe Systems. 

Organism Tested



Time (Hours)

Special Reaction

Bacteroides fragilis **



24 h


Prevotella melaninogenica **



48 h


Fusobacterium nucleatum **



24 h


Clostridium perfringens **



24 h

Double Zone of hemolysis

Peptostreptococcus anaerobius **



24 h


Staphylococcus aureus   or    Enterococcus faecalis

25923     29212


24 h


Escherichia coli



24 h


Proteus mirabilis



24 h


Propionibacterium acnes   or  Clostridium difficile

6919      9686


24 – 48 h

24 h


** 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.



Growth in Hours

Expected Results

B. fragilis

24 hrs


C. perfringens

24 hrs

Double zone of b -hemolysis

P. anaerobius

24 hrs


E. faecalis

24 hrs



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



1.        Dowell, V. R., Jr. and T. M. Hawkins.  1974Laboratory 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.  1977Anaerobe Laboratory ManualVirginia 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 ManualStar Publishing Co., Belmont, CA 94002.


4.        NCCLSQuality 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 BacteriologyStar Publishing Co., Belmont, CA 94002.