Anaerobe Systems PRAS Crystal-Violet
                 Erythromycin (CVE) AS-647

Formula

Pancreatic Digest of Caesin, 10.0 g

Yeast Extract, 5.0 g

Sodium Chloride, 5.0 g

Dextrose, 2.0 g

Tryptophan, 0.2 g

Agar, 15.0 g

Crystal Violet, 0.005 g

Erythromycin, 0.004 g

Sheep Blood, 50.0 ml

Distilled Water, 1000.0 ml

 

Final pH 7.2 +/- 0.3 at 25^oC.

Final weight 16.0 g +/- 1.6 for AS-647.

                                                                                                               

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 air (oxygen) during collection, transport and processing.  Consult appropriate references for detailed instructions concerning collection and transport of anaerobes.

Methods for Use:  CVE 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 35-37^oC 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 (1, 2, 3, 4, 5, 6).

Materials Required But Not Provided

Standard microbiological supplies and equipment such as loops, saline blanks, slides, staining supplies, microscope, incinerator / autoclave, incubators, anaerobic chamber, other culture media and serological and biochemical reagents.

 

Limitations

CVE will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  It is recommended that a non-selective medium such as Anaerobic Brucella Blood Agar also be inoculated from clinical specimens to assure growth of all species present.  Consult reference materials for additional information.

 

Quality Control

CVE, if used properly, should support good growth of Fusobacterium nucleatum found in clinical infections.

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

Organism Tested	ATCC #	Results	Time (Hours)	Special Reaction
Fusobacterium nucleatum	25586	Growth	24 hrs	Blue pigment
Actinobacillus actinomycetemcomitans	29522	Growth	24 – 48 hrs
Porphyromonas asaccharolytica	25260	No Growth
Bacteroides fragilis	25285	Inhibited
Peptostreptococcus anaerobius	27337	No Growth
Clostridium perfringens	13124	Inhibited
Enterococcus faecalis	29212	Inhibited
Actinomyces viscosus	43146	No Growth
Fusobacterium necrophorum	25286	Growth	24 hrs	Blue pigment
Propionibacterium acnes	6919	No Growth
Clostridium difficile	9689	No Growth

 

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/inhibitory capacity of this medium is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth/inhibition.

 

Organism	Expected Growth	Special Reaction
F. nucleatum	24 hours	Blue pigment
A. actinomycetemcomitans	48 hours	-
A. viscosus	Inhibited
P. asaccharolytica	Inhibited

 

Physical Appearance:  CVE should appear opaque and purple 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.        Engelkirk, P. G., Duben-Engelkirk, J., Dowell, V. R.  1992.  Principles of Practices of Clinical Anaerobic Bacteriology. Star Publishing Co., Belmont, CA 94002.

 

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

 

4.        Jousimies-Somer, 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.

 

5.        Walker, V. L., Ratliff, D., Muller, D., Mandell, R. and S. Socransky.  Medium for Selective Isolation of Fusobacterium nucleatum from Human Periodontal Pockets.  Journal of Clinical Microbiology.  Dec., 1979 Vol. 10.

 

6.        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.