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Journal of Clinical Microbiology, June 2009, p. 1888-1891, Vol. 47, No. 6
0095-1137/09/$08.00+0     doi:10.1128/JCM.00506-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay{triangledown}

Erik Munson,1,2* Timothy Block,1,{dagger} Janet T. Voegeli,1 Jeanne E. Hryciuk,1 and Ronald F. Schell3,4

Wheaton Franciscan and Midwest Clinical Laboratories, Wauwatosa, Wisconsin 53226,1 College of Health Sciences, University of Wisconsin—Milwaukee, Milwaukee, Wisconsin 53201,2 Wisconsin State Laboratory of Hygiene,3 Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, Wisconsin 537064

Received 12 March 2009/ Accepted 14 April 2009

The expense inherent to molecular diagnostics may be an overriding concern for a variety of clinical laboratories in the development of PCR-based methicillin-resistant Staphylococcus aureus (MRSA) active surveillance programs. BD GeneOhm MRSA assay master mix was reconstituted, aliquoted into SmartCycler tubes in 25-µl volumes, and frozen at –70°C. One hundred percent of archival nasal swab lysates yielded the expected PCR results when incubated in master mix frozen for 1, 2, 3, and 4 weeks. A 98.8% concordance of the final result was observed upon prospective PCR analysis of 320 clinical lysates utilizing freshly reconstituted master mix and 2-week-frozen master mix. Initial unresolved rates generated by frozen master mix and freshly reconstituted master mix differed by 1.6% (P = 0.16). Of 50 MRSA-positive lysates, the titers of 32 (64%) were determined to the same value upon initial tandem frozen master mix and freshly reconstituted master mix utilization; the titers of an additional 14 were determined to the same value upon repeat testing. Frozen master mix maintains potency for at least 4 weeks, facilitating detection of MRSA from nasal swab lysates, and may decrease the amount of unused reagent up to an average of 33%.

* Corresponding author. Mailing address: Wheaton Franciscan Laboratory, 11020 West Plank Court, Suite 100, Wauwatosa, WI 53226. Phone: (414) 256-1479. Fax: (414) 256-5566. E-mail: Erik.Munson{at}wfhc.org

{triangledown} Published ahead of print on 22 April 2009.

{dagger} Present address: St. Joseph's Hospital, West Bend, WI 53095.

Journal of Clinical Microbiology, June 2009, p. 1888-1891, Vol. 47, No. 6
0095-1137/09/$08.00+0     doi:10.1128/JCM.00506-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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