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U
IX PROCEDURE
Material Provided: Mueller Hinton Broth
Materials Required But Not Provided: Ancillary culture media, reagents, quality control organisms and laboratory equipment as
required.
Test Procedure: Observe aseptic techniques and established precautions against microbiological hazards throughout all
procedures.
Organisms to be subcultured must first be isolated in pure culture on an appropriate solid medium. Transfer growth from the
isolation medium to Mueller Hinton Broth using standard bacteriologic techniques.
5-7
For enrichment purposes, inoculate the specimen onto primary media and then into the broth according to recommended
procedures.
Incubate tubes and bottles at 35C under conditions appropriate for the organism being cultured.
User Quality Control: See "Quality Control Procedures."
Quality control requirements must be performed in accordance with applicable local, state and/or federal regulations or
accreditation requirements and your laboratorys standard Quality Control procedures. It is recommended that the user refer to
pertinent CLSI guidance and CLIA regulations for appropriate Quality Control practices.
X RESULTS
Growth in broth media is indicated by the presence of turbidity compared with an uninoculated control.
XI LIMITATIONS OF THE PROCEDURE
Enrichment broths should not be used as the sole isolation medium. They are to be used in conjunction with selective and
nonselective plating media to increase the probability of isolating pathogens, especially when they may be present in small
numbers.
For identification, organisms must be in pure culture. Morphological, biochemical and/or serological tests should be performed
for final identification. Consult appropriate texts for further information.
4-7
XII PERFORMANCE CHARACTERISTICS
Prior to release, all lots of Mueller Hinton Broth are tested for performance characteristics. Representative samples of the lot
are tested with cell suspensions of Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922 and Staphylococcus aureus
ATCC 25923, diluted in normal saline to yield 10
3
to 10
4
CFU per tube of broth. Tubes are incubated with loose caps at 35 2C
for one day in an aerobic atmosphere. Growth is observed with all organisms.
XIII AVAILABILITY
Cat. No. Description
296164 BBL Mueller Hinton Broth, 2 mL, Ctn. of 100 size K tubes
XIV REFERENCES
1. Mueller, J.H., and J. Hinton. 1941. A protein-free medium for primary isolation of he gonococcus and meningooccus. Proc.
Soc. Exp. Biol. Med. 48:330-333.
2. Clinical and Laboratory Standards Institute. 2009. Approved standard M7-A8. Methods for dilution antimicrobial susceptibility
tests for bacteria that grow aerobically, 8th ed. CLSI, Wayne, Pa.
3. Clinical and Laboratory Standards Institute. 2009. Approved standard M2-A10. Performance standards for
antimicrobial disk susceptibility tests, 10th ed. CLSI, Wayne, Pa.
4. Washington, J.A. (ed.). 1985. Laboratory procedures in clinical microbiology, 2nd ed. Springer-Verlag, New York.
5. Forbes, B.A., D.F. Sahm, and A.S. Weissfeld. 1998. Bailey & Scotts diagnostic microbiology, 10th ed. Mosby, Inc., St. Louis.
6. Murray, P.R., E.J. Baron, M.A. Pfaller, F.C. Tenover, and R.H. Yolken (ed.). 1995. Manual of clinical microbiology, 6th ed.
American Society for Microbiology, Washington, D.C.
7. Holt, J.G., N.R. Krieg, P.H.A. Sneath, J.T. Staley, and S.T. Williams (ed.). 1994. Bergeys Manual of determinative bacteriology,
9th ed. Williams & Wilkins, Baltimore.
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