Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay
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 SmartC...
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| Veröffentlicht in: | Journal of Clinical Microbiology 2009-06, Vol.47 (6), p.1888-1891 |
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| container_title | Journal of Clinical Microbiology |
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| creator | Munson, Erik Block, Timothy Voegeli, Janet T Hryciuk, Jeanne E Schell, Ronald F |
| description | 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%. |
| doi_str_mv | 10.1128/JCM.00506-09 |
| format | Article |
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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%.</description><identifier>ISSN: 0095-1137</identifier><identifier>EISSN: 1098-660X</identifier><identifier>DOI: 10.1128/JCM.00506-09</identifier><identifier>PMID: 19386849</identifier><identifier>CODEN: JCMIDW</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Bacteriology ; Biological and medical sciences ; Carrier State - diagnosis ; Freezing ; Fundamental and applied biological sciences. Psychology ; Humans ; Mass Screening - economics ; Mass Screening - methods ; Methicillin-Resistant Staphylococcus aureus - isolation & purification ; Microbiology ; Miscellaneous ; Molecular Diagnostic Techniques - economics ; Molecular Diagnostic Techniques - methods ; Polymerase Chain Reaction - economics ; Polymerase Chain Reaction - methods ; Reagent Kits, Diagnostic - economics ; Sensitivity and Specificity ; Staphylococcal Infections - diagnosis ; Staphylococcus aureus</subject><ispartof>Journal of Clinical Microbiology, 2009-06, Vol.47 (6), p.1888-1891</ispartof><rights>2009 INIST-CNRS</rights><rights>Copyright © 2009, American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c493t-7335f15d7d1f80a98fb2f96c37719ef9b4fec1717aba4ae0a1e26602d98879f53</citedby><cites>FETCH-LOGICAL-c493t-7335f15d7d1f80a98fb2f96c37719ef9b4fec1717aba4ae0a1e26602d98879f53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2691094/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2691094/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,3189,3190,27928,27929,53795,53797</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21551090$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19386849$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Munson, Erik</creatorcontrib><creatorcontrib>Block, Timothy</creatorcontrib><creatorcontrib>Voegeli, Janet T</creatorcontrib><creatorcontrib>Hryciuk, Jeanne E</creatorcontrib><creatorcontrib>Schell, Ronald F</creatorcontrib><title>Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay</title><title>Journal of Clinical Microbiology</title><addtitle>J Clin Microbiol</addtitle><description>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%.</description><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Carrier State - diagnosis</subject><subject>Freezing</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Mass Screening - economics</subject><subject>Mass Screening - methods</subject><subject>Methicillin-Resistant Staphylococcus aureus - isolation & purification</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Molecular Diagnostic Techniques - economics</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Polymerase Chain Reaction - economics</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Reagent Kits, Diagnostic - economics</subject><subject>Sensitivity and Specificity</subject><subject>Staphylococcal Infections - diagnosis</subject><subject>Staphylococcus aureus</subject><issn>0095-1137</issn><issn>1098-660X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtvEzEURkcIRENhxxrMAlZMscfj1wapGvUBagRqqcTOunHsxNXMONieQvj1uCQqsGLju_DRp_vdU1XPCT4ipJHvPnbzI4wZ5jVWD6oZwUrWnOOvD6sZxorVhFBxUD1J6QZj0raMPa4OiKKSy1bNqtSFlOsT56zJ_tai0xh-2hHNIWUb0dz_QPOw9M4byD6MKDgEqAvDYKPx0KO5zWtvfN_7sb60yacMY0ZXGTbrbR9MMGZKCKZoy_jcXaLjlGD7tHrkoE_22X4eVtenJ1-68_ri09mH7viiNq2iuRaUMkfYUiyJkxiUdIvGKW6oEERZpxZtWZoIImABLVgMxDaleLNUUgrlGD2s3u9yN9NisEtjxxyh15voB4hbHcDrf39Gv9arcKsbrsoZ2xLwZh8Qw7fJpqwHn4ztexhtmJLmghJKmPov2BRRgjNewLc70MSQUrTufhuC9Z1OXXTq3zo1vst98XeDP_DeXwFe7wFIBnoXYTQ-3XMNYaw0wYV7tePWfrX-7qPVkAZ9YwbdCs01kVIW5uWOcRA0rGLJub5qMKGYcMqb8vwCug2-Kw</recordid><startdate>20090601</startdate><enddate>20090601</enddate><creator>Munson, Erik</creator><creator>Block, Timothy</creator><creator>Voegeli, Janet T</creator><creator>Hryciuk, Jeanne E</creator><creator>Schell, Ronald F</creator><general>American Society for Microbiology</general><general>American Society for Microbiology (ASM)</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>C1K</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20090601</creationdate><title>Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay</title><author>Munson, Erik ; Block, Timothy ; Voegeli, Janet T ; Hryciuk, Jeanne E ; Schell, Ronald F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c493t-7335f15d7d1f80a98fb2f96c37719ef9b4fec1717aba4ae0a1e26602d98879f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Carrier State - diagnosis</topic><topic>Freezing</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Mass Screening - economics</topic><topic>Mass Screening - methods</topic><topic>Methicillin-Resistant Staphylococcus aureus - isolation & purification</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Molecular Diagnostic Techniques - economics</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Polymerase Chain Reaction - economics</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Reagent Kits, Diagnostic - economics</topic><topic>Sensitivity and Specificity</topic><topic>Staphylococcal Infections - diagnosis</topic><topic>Staphylococcus aureus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Munson, Erik</creatorcontrib><creatorcontrib>Block, Timothy</creatorcontrib><creatorcontrib>Voegeli, Janet T</creatorcontrib><creatorcontrib>Hryciuk, Jeanne E</creatorcontrib><creatorcontrib>Schell, Ronald F</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Clinical Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Munson, Erik</au><au>Block, Timothy</au><au>Voegeli, Janet T</au><au>Hryciuk, Jeanne E</au><au>Schell, Ronald F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay</atitle><jtitle>Journal of Clinical Microbiology</jtitle><addtitle>J Clin Microbiol</addtitle><date>2009-06-01</date><risdate>2009</risdate><volume>47</volume><issue>6</issue><spage>1888</spage><epage>1891</epage><pages>1888-1891</pages><issn>0095-1137</issn><eissn>1098-660X</eissn><coden>JCMIDW</coden><abstract>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. 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| source | American Society for Microbiology; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central |
| subjects | Bacteriology Biological and medical sciences Carrier State - diagnosis Freezing Fundamental and applied biological sciences. Psychology Humans Mass Screening - economics Mass Screening - methods Methicillin-Resistant Staphylococcus aureus - isolation & purification Microbiology Miscellaneous Molecular Diagnostic Techniques - economics Molecular Diagnostic Techniques - methods Polymerase Chain Reaction - economics Polymerase Chain Reaction - methods Reagent Kits, Diagnostic - economics Sensitivity and Specificity Staphylococcal Infections - diagnosis Staphylococcus aureus |
| title | Cost-Effective Frozen Master Mix Modification of a Commercial Methicillin-Resistant Staphylococcus aureus PCR Assay |
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