Quality evaluation of LC‐MS/MS‐based E. coli H antigen typing (MS‐H) through label‐free quantitative data analysis in a clinical sample setup. Issue 11 (30th October 2014)

• Record Type: Journal Article
• Authors: Cheng, Keding; Sloan, Angela; McCorrister, Stuart; Peterson, Lorea; Chui, Huixia; Drebot, Mike; Nadon, Celine; Knox, J. David; Wang, Gehua; Everett, Allen; Ignjatovic, Vera
• Is Part Of: Proteomics. Volume 8:Issue 11/12(2014)
• Page Start: 963
• Page End: 970
• Publication Date: 2014-10-30
• Subjects: Proteomics -- Periodicals; 572.605
• DOI: 10.1002/prca.201400019
• Languages: English
• ISSNs: 1862-8346

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="prca1581-sec-0010" sec-type="section"> <title>Purpose</title> <p>The need for rapid and accurate H typing is evident during <italic>Escherichia coli</italic> outbreak situations. This study explores the transition of MS‐H, a method originally developed for rapid H antigen typing of <italic>E. coli</italic> using LC‐MS/MS of flagella digest of reference strains and some clinical strains, to <italic>E. coli</italic> isolates in clinical scenario through quantitative analysis and method validation.</p> </sec> <sec id="prca1581-sec-0020" sec-type="section"> <title>Experimental design</title> <p>Motile and nonmotile strains were examined in batches to simulate clinical sample scenario. Various LC‐MS/MS batch run procedures and MS‐H typing rules were compared and summarized through quantitative analysis of MS‐H data output for a standard method development.</p> </sec> <sec id="prca1581-sec-0030" sec-type="section"> <title>Results</title> <p>Label‐free quantitative data analysis of MS‐H typing was proven very useful for examining the quality of MS‐H result and the effects of some sample carryovers from motile <italic>E. coli</italic> isolates. Based on this, a refined procedure and protein identification rule specific for clinical MS‐H typing was established and validated.</p> </sec> <sec id="prca1581-sec-0040" sec-type="section"> <title>Conclusions and clinical relevance</title> <p>With LC‐MS/MS batch run procedure and database search parameter unique for <italic>E. coli</italic> MS‐H typing, the standard procedure maintained high accuracy and specificity in clinical situations, and its potential to be used in a clinical setting was clearly established.</p> </sec> </abstract>