{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T00:33:30Z","timestamp":1777682010231,"version":"3.51.4"},"reference-count":16,"publisher":"SAGE Publications","issue":"2","license":[{"start":{"date-parts":[[2013,5,15]],"date-time":"2013-05-15T00:00:00Z","timestamp":1368576000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2013,5,15]],"date-time":"2013-05-15T00:00:00Z","timestamp":1368576000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Biosecurity and Bioterrorism: Biodefense Strategy, Practice, and Science"],"published-print":{"date-parts":[[2013,6,1]]},"abstract":"<jats:p>Molecular bioforensic research is dependent on rapid and sensitive methods such as real-time PCR (qPCR) for the identification of microorganisms. The use of synthetic positive control templates containing small modifications outside the primer and probe regions is essential to ensure all aspects of the assay are functioning properly, including the primers and probes. However, a typical qPCR or reverse transcriptase qPCR (qRT-PCR) assay is limited in differentiating products generated from positive controls and biological samples because the fluorescent probe signals generated from each type of amplicon are indistinguishable. Additional methods used to differentiate amplicons, including melt curves, secondary probes, and amplicon sequencing, require significant time to implement and validate and present technical challenges that limit their use for microbial forensic applications. To solve this problem, we have developed a novel application of electrospray ionization mass spectrometry (ESI-MS) to rapidly differentiate qPCR amplicons generated with positive biological samples from those generated with synthetic positive controls. The method has sensitivity equivalent to qPCR and supports the confident and timely determination of the presence of a biothreat agent that is crucial for policymakers and law enforcement. Additionally, it eliminates the need for time-consuming methods to confirm qPCR results, including development and validation of secondary probes or sequencing of small amplicons. In this study, we demonstrate the effectiveness of this approach with microbial forensic qPCR assays targeting multiple biodefense agents (bacterial, viral, and toxin) for the ability to rapidly discriminate between a positive control and a positive sample.<\/jats:p>","DOI":"10.1089\/bsp.2012.0062","type":"journal-article","created":{"date-parts":[[2013,5,15]],"date-time":"2013-05-15T09:58:00Z","timestamp":1368611880000},"page":"107-117","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":2,"title":["Differentiating Microbial Forensic qPCR Target and Control Products by Electrospray Ionization Mass Spectrometry"],"prefix":"10.1177","volume":"11","author":[{"given":"S. Timothy","family":"Motley","sequence":"first","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cassie L.","family":"Redden","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristin A.","family":"Sannes-Lowery","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark W.","family":"Eshoo","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steven A.","family":"Hofstadler","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James P.","family":"Burans","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. J.","family":"Rosovitz","sequence":"additional","affiliation":[{"name":"S. Timothy Motley, MS, is Principal Scientist, New Technology Development; Mark W. Eshoo, PhD, is Director, New Technology Development; Kristin A. Sannes-Lowery, PhD, is Director of Commercial Laboratory Operations; and Steven A. Hofstadler, PhD, is Divisional Vice President, Research and Development; all at Ibis Biosciences, Inc., An Abbott Company, Carlsbad, California. Cassie L. Redden, BS, was, at the time this article was written, Research Associate; James P. Burans, PhD, is Director; and M. J. Rosovitz, PhD, is Principal Investigator; all at the National Bioforensic Analysis Center, National Biodefense Analysis and Countermeasures Center, Frederick, Maryland."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2013,5,15]]},"reference":[{"issue":"10","key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"3446","DOI":"10.1128\/JCM.39.10.3446-3451.2001","article-title":"Sensitive and specific method for rapid identification of Streptococcus pneumoniae using real-time fluorescence PCR","volume":"39","author":"McAvin JC","year":"2001","unstructured":"McAvin JC, , Reilly PA, , Roudabush RM et al. Sensitive and specific method for rapid identification of Streptococcus pneumoniae using real-time fluorescence PCR. J Clin Microbiol, 2001; 39,10:3446\u20133451.","journal-title":"J Clin Microbiol"},{"issue":"1","key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"S39","DOI":"10.1016\/S1386-6532(02)00032-X","article-title":"Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens","volume":"25","author":"Verstrepen WA","year":"2002","unstructured":"Verstrepen WA, , Bruynseels P, , Mertens AH. Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens. J Clin Virol, 2002; 25,Suppl 1:S39\u2013S43.","journal-title":"J Clin Virol"},{"issue":"9","key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"3985","DOI":"10.1128\/AEM.67.9.3985-3993.2001","article-title":"Detection of legionellae in hospital water samples by quantitative real-time LightCycler PCR","volume":"67","author":"Wellinghausen N","year":"2001","unstructured":"Wellinghausen N, , Frost C, , Marre R. Detection of legionellae in hospital water samples by quantitative real-time LightCycler PCR. Appl Environ Microbiol, 2001; 67,9:3985\u20133993.","journal-title":"Appl Environ Microbiol"},{"issue":"3","key":"e_1_3_2_5_2","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S1386-6532(03)00072-6","article-title":"Co-detection and discrimination of six human herpesviruses by multiplex PCR-ELAHA","volume":"28","author":"Mackay IM","year":"2003","unstructured":"Mackay IM, , Gardam T, , Arden KE et al. Co-detection and discrimination of six human herpesviruses by multiplex PCR-ELAHA. J Clin Virol, 2003; 28,3:291\u2013302.","journal-title":"J Clin Virol"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0409920102"},{"issue":"7","key":"e_1_3_2_7_2","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/nrmicro1918","article-title":"Ibis T5000: a universal biosensor approach for microbiology","volume":"6","author":"Ecker DJ","year":"2008","unstructured":"Ecker DJ, , Sampath R, , Massire C et al. Ibis T5000: a universal biosensor approach for microbiology. Nat Rev Microbiol, 2008; 6,7:553\u2013558.","journal-title":"Nat Rev Microbiol"},{"issue":"3","key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1128\/JCM.01578-10","article-title":"Simultaneous identification of mycobacterial isolates to the species level and determination of tuberculosis drug resistance by PCR followed by electrospray ionization mass spectrometry","volume":"49","author":"Massire C","year":"2011","unstructured":"Massire C, , Ivy CA, , Lovari R et al. Simultaneous identification of mycobacterial isolates to the species level and determination of tuberculosis drug resistance by PCR followed by electrospray ionization mass spectrometry. J Clin Microbiol, 2011; 49,3:908\u2013917.","journal-title":"J Clin Microbiol"},{"issue":"2","key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1111\/j.1365-2672.2009.04215.x","article-title":"Development of real-time PCR tests for detecting botulinum neurotoxins A, B, E, F producing Clostridium botulinum, Clostridium baratii and Clostridium butyricum","volume":"107","author":"Fach P","year":"2009","unstructured":"Fach P, , Micheau P, , Mazuet C, , Perelle S, , Popoff M. Development of real-time PCR tests for detecting botulinum neurotoxins A, B, E, F producing Clostridium botulinum, Clostridium baratii and Clostridium butyricum. J Appl Microbiol, 2009; 107,2:465\u2013473.","journal-title":"J Appl Microbiol"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2006.06.002"},{"key":"e_1_3_2_11_2","first-page":"174","article-title":"Development of a duplex quantitative real-time PCR assay for the detection of tick-borne spotted fever group rickettsiae and Rickettsia rickettsii","volume":"78","author":"Jiang J","year":"2005","unstructured":"Jiang J, , Blair P, , Olson J, , Stromdahl E, , Richards A. Development of a duplex quantitative real-time PCR assay for the detection of tick-borne spotted fever group rickettsiae and Rickettsia rickettsii. Int Rev Armed Forces Med Serv, 2005; 78:174\u2013179.","journal-title":"Int Rev Armed Forces Med Serv"},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1111\/j.1749-6632.2003.tb07380.x","article-title":"Real-time PCR duplex assay for Rickettsia prowazekii and Borrelia recurrentis","volume":"990","author":"Jiang J","year":"2003","unstructured":"Jiang J, , Temenak JJ, , Richards AL. Real-time PCR duplex assay for Rickettsia prowazekii and Borrelia recurrentis. Ann NY Acad Sci, 2003; 990:302\u2013310.","journal-title":"Ann NY Acad Sci"},{"issue":"2","key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.jviromet.2004.05.018","article-title":"Specific detection of Nipah virus using real-time RT-PCR (TaqMan)","volume":"120","author":"Guillaume V","year":"2004","unstructured":"Guillaume V, , Lefeuvre A, , Faure C et al. Specific detection of Nipah virus using real-time RT-PCR (TaqMan) J Virol Methods, 2004; 120,2:229\u2013237.","journal-title":"J Virol Methods"},{"issue":"1","key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0166-0934(01)00354-8","article-title":"Development of a fluorogenic RT-PCR assay (TaqMan) for the detection of Hendra virus","volume":"98","author":"Smith IL","year":"2001","unstructured":"Smith IL, , Halpin K, , Warrilow D, , Smith GA. Development of a fluorogenic RT-PCR assay (TaqMan) for the detection of Hendra virus. J Virol Methods, 2001; 98,1:33\u201340.","journal-title":"J Virol Methods"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1103.040629"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.5.3.312"},{"issue":"2","key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.diagmicrobio.2010.10.010","article-title":"Reverse transcription polymerase chain reaction and electrospray ionization mass spectrometry for identifying acute viral upper respiratory tract infections","volume":"69","author":"Chen KF","year":"2011","unstructured":"Chen KF, , Blyn L, , Rothman RE et al. Reverse transcription polymerase chain reaction and electrospray ionization mass spectrometry for identifying acute viral upper respiratory tract infections. Diagn Microbiol Infect Dis, 2011; 69,2:179\u2013186.","journal-title":"Diagn Microbiol Infect Dis"}],"container-title":["Biosecurity and Bioterrorism: Biodefense Strategy, Practice, and Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1089\/bsp.2012.0062","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/bsp.2012.0062","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/bsp.2012.0062","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T08:41:37Z","timestamp":1777452097000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/10.1089\/bsp.2012.0062"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,5,15]]},"references-count":16,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2013,6,1]]}},"alternative-id":["10.1089\/bsp.2012.0062"],"URL":"https:\/\/doi.org\/10.1089\/bsp.2012.0062","relation":{},"ISSN":["1538-7135","1557-850X"],"issn-type":[{"value":"1538-7135","type":"print"},{"value":"1557-850X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,5,15]]}}}