DOC # CS-100365 REV 0

Product Insert PRAS DILUTION BLANK Products: AS-907

PRAS Dilution Blanks, 7 mL

10 tubes / pkg

AS-908

PRAS Dilution Blanks, 9 mL

10 tubes / pkg

AS-910

PRAS Dilution Blanks, 10 mL

10 tubes / pkg

AS-917

PRAS Dilution Blanks, 3 mL

10 tubes / pkg

AS-918

PRAS Dilution Blanks, 18 mL

10 tubes / pkg

Intended Use PRAS (Pre-reduced Anaerobically Sterilized) Dilution Blanks are a mineral salt base liquid with reducing agents designed as a holding medium for maintaining viability of microorganisms, especially anaerobic bacteria, in order to determine the number of viable organisms present in a sample.

Summary Bacterial cells are often diluted from their original dense concentration to a sparse concentration to become suitable for use in microscope, quantitation, genetic analysis or metabolic studies. PRAS Dilution Blanks contain buffered mineral salts with sodium thioglycolate and L-cysteine added to provide a reduced environment. This combination has been prepared to provide an environment which maintains viability of most microorganisms without significant multiplication. PRAS Dilution Blanks are designed to meet the stringent viability requirements of obligate anaerobes. The blanks are supplied in tubes with a screw cap and/or rubber septa (Hungate caps), which allow for direct injection of aspirated clinical material. This medium is prepared, dispensed, stored and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

Formula (g/L) Magnesium Sulfate, Heptahydrate Potassium Phosphate, Monobasic Potassium Chloride Sodium Phosphate, Dibasic Sodium Chloride Sodium Thioglycollate L-Cysteine (25.0% solution) DI Water

0.1 g 0.2 g 0.2 g 1.15 g 3.0 g 1.0 g 2.0 mL 1.0 L

Final pH 7.5 +/- 0.5 at 25oC. Final volume 7.0 mL +/- 0.7 for AS-907. Final volume 9.0 mL +/- 0.9 for AS-908. Final volume 10.0 mL +/- 1.0 for AS-910. Final volume 3.0 mL +/- 0.3 for AS-917. Final volume 18.0 mL +/- 1.8 for AS-918.

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 packaging until use. Avoid overheating or freezing. Do not use medium if there are signs of deterioration (discoloration or evaporation) 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: 1 year from date of manufacture.

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Anaerobe Systems 15906 Concord Circle Morgan Hill, CA 95037 408 782 7557 Fax 408 782 3031 http://www.anaerobesystems.com

DOC # CS-100365 REV 0

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: PRAS Dilution Blanks should be inoculated with a 0.5 McFarland of bacterial suspension, by aseptic and anaerobic technique. After completion of serial ten-fold dilutions that produce statistically valid plate counts, immediately place on Brucella medium. For syringe application, the rubber septum in the cap should be disinfected and the fluid specimen directly injected into the tube at a slow rate. For either procedure type, the inoculated tubes should be kept at room temperature and processed as soon as possible. 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, pipettes, slides, staining supplies, microscope, incinerator/autoclave, incubators, anaerobic chamber or anaerobic jars, disinfectant, other culture media and serological and biochemical reagents.

Limitations PRAS Dilution Blanks are designed to be a holding medium to maintain viability of microorganisms contained in a specimen. Other media will be required for isolation and identification of clinical isolates. Specimens should be transported to the laboratory and processed promptly since delayed processing may result in overgrowth by one organism in specimens from polymicrobic infections. Consult reference materials for additional information.

Quality Control If used properly, this medium should maintain the viability of microorganisms within clinical material in order to determine the number of viable organisms. Organisms are serially diluted and recovered on BRU agar. The following organisms are routinely used for quality assurance performance testing at Anaerobe Systems. Organism Tested ATCC # Results Time (Hours) Bacteroides fragilis 25285 Growth 24 Prevotella melaninogenica 25845 Growth 24 – 48 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 holding capacity of this media is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth. Organism Expected Growth B. fragilis 24 hrs P. melaninogenica 24-48 hrs Physical Appearance: PRAS Dilution Blanks should appear as a clear liquid within a 16mm x 100mm glass tube (for AS-907, AS-908, AS-910, & AS-917) or a 20mm x 113mm glass tube (AS-918).

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.

Dowell, V. R., Jr. and G. L. Lombard. 1977. Presumptive Identification of Anaerobic Non-sporeforming Gram-negative Bacilli. USDHEW, CDC. Atlanta, GA 30333.

3.

Dowell, V. R., G. L. Lombard, S. F. Thompson and A. Y. Armfield. 1977. Media for the Isolation, Characterization and Identification of Obligately Anaerobic Bacteria. USDHEW, CDC. Atlanta, GA 30333.

4.

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

5.

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

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.

7.

Gerhardt, P., Murray, R. G. E., Costilow, R. N., Nester, E. W., Wood, W. A, Krieg, N. R. and Philips, G. B. 1981. Manual of Methods for General Bacteriology. ASM Washington DC 20006.

Revision Date: 8/31/15 Page 2 of 2

Anaerobe Systems 15906 Concord Circle Morgan Hill, CA 95037 408 782 7557 Fax 408 782 3031 http://www.anaerobesystems.com