TRYPTIC SOY AGAR (TSA) W/ 5% SHEEP BLOOD

Technical Data #1993a / 2013.06.16

Tryptic Soy Agar w/ 5% sheep blood is a general purpose culture medium for cultivation and isolation of fastidious or nonfastidious microorganisms. The addition of blood permits the interpretation of hemolytic activities.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

15,0

Soy Peptone

5,0

Sodium Chloride

5,0

Agar

15,0

Defibrinated Sheep Blood

50 ml

pH 7.3 +/- 0,2 25 C

This approximate formula may be adjusted and/or enriched to obtain best results.


PRECAUTIONS
This medium is for IN VITRO diagnostic use only.


STORAGE
Store prepared media at 2-8 C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25 C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 40 g of dehydrated media in 950 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121 C for 15 minutes. Cool to 45-50 C and aseptically, add 50 ml of Defibrinated Sheep Blood (4245). Mix gently and dispense into sterile Petri dishes or sterile culture tubes.


PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

  1. Directly, inoculate plate or tubes as soon as possible with specimen and streak for isolation. Several stabs are recommended to be done in blood plates.
  2. Incubate at 35 C under CO2 conditions for 24-48 hours.
  3. Examine for growth and hemolysis.


QUALITY CONTROL
 

Results after 24 hrs at 35C W CO2
Organisms

ATCC

Growth

Hemolysis

Streptococcus pyogenes

19615

+

beta

Streptococcus pneumoniae

6305

+

alpha

Staphylococcus aureus

25923

+

 
Escherichia coli

25922

+

 

Camp test results after 24 hrs at 35C

Organisms

ATCC

Growth

Staphylococcus aureus

33682

 

Streptococcus agalactiae

12386

positive,
arrowhead area of clearing

Streptococcus pyogenes

19615

negative,
no arrowhead formation


LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.


REFERENCES

  1. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American Society for Microbiology.
  2. N.C.C.L.S. 1990 Quality Assurance for Commercially Prepared Microbiological Culture Media. Approved Standard. Vol.10, No.14 NCCLS Document M22-A.
  3. Mac Faddin, Jean. F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol. 1. Baltimore, MD.: Williams & Wilkins.
  4. Clesceri, L.S., A.E. Greenberg, and R.R. Trussell. 1989 Standard Methods for the Examination of Water and Wastewater. 17th ed. American Public Health Association, Washington, D.C.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-5106
     
Prepared media    
Plates, 100x15 mm 10/pkg 1993
Mono-Plates, 100x15 mm 10/pkg 1993M
Tubes, 16x125 mm 10/pkg 2635
     
Defibrinated Sheep Blood 50 ml 4245

 


 





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