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Application of MALDI-TOF Mass Spectrometry for Rapid Pathogen Identification in an Era of ‘One Medicine-One Health’ Microbiology T

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Application of MALDI-TOF Mass Spectrometry for Rapid Pathogen Identification in an Era of ‘One Medicine-One Health’ Microbiology T Poster P-2331 ECCMID, 2012 Application of MALDI-TOF Mass Spectrometry for Rapid Pathogen Identification in an Era of ‘One Medicine-One Health’ Microbiology T. Fritsche1,3, B. Olson2, C. Pike1, M. Stemper1,3, F. Moore1, J. Meece2,3, S. Shukla2,3, T. Novicki1 Marshfield Clinic, Marshfield, WI, USA1. Marshfield Clinic Res. Fndn., Marshfield, WI ,USA2. Univ. of Wisconsin, La Crosse, WI, USA3 REVISED ABSTRACT* INTRODUCTION OBJECTIVES: To evaluate the utility of MALDI-TOF mass The majority of human bacterial and fungal pathogens (up to 60% by some assessments) also produce disease in Table 2. Comparison (%) of genus- and species-level spectrometry (MS) as a single platform for pathogen various animal species. The remainder comprise a variety of species that are more specific pathogens for the particular identifications using MBT and Standard Lab Methods (SLM). identification (ID) in a regional laboratory practice host, and/or that only cross species barriers occasionally (zoonoses). Differences in laboratory practices between routinely performing analyses on human- and animal- human and veterinary testing laboratories relate to the differences in disease-producing agents, identification schema, Human Source Animal Source Overall source clinical specimens. Integration of signature- different antimicrobial agents and different guidance documents. SLM Genus Level ID 94.1 88.2 91.2 based proteomic analyses into medical and veterinary SLM Species Level ID 70.0 51.8 60.9 laboratories may enhance ID performance, improve Despite such differences, the increasing recognition that many of our core pathogens increasingly display similar MBT Genus Level ID 98.8 95.4 97.1 clinical service and epidemiologic research, and antimicrobial resistance phenotypes, and that we readily share these pathogens with our companion and food-source promote cross-disciplinary collaboration promulgated animals, behooves us to consider simultaneously the emergence and significance of these ‘shared’ pathogens in both MBT Species Level ID 94.4 87.2 90.8 by the ‘One Medicine-One Health’ concept (Veterinaria environments. Such recognition of the need for greater integration of human and animal health concerns has been the Italiana 45[1]; 2009). Here we demonstrate that MS can impetus for some years in the promulgation of the ‘One Medicine-One Health’ concept by the medical, veterinary and be as rapid, accurate and cost-effective when testing public health professions (Kaplan, 2009; Kahn, 2009). The lack of opportunity to capitalize on this concept is a practical isolates of veterinary importance as when testing matter; whether the human and animal laboratory testing communities are prepared to assist each other with this CONCLUSIONS human-source isolates in the same laboratory setting. initiative remains to be determined, but the benefits are considerable (Cardiff, 2008). •Use of mass spectrometry-generated signature-based proteomic analyses for pathogen identification is rapidly METHODS: Clinically significant pathogens (n=1,502) Technology is one area where medical and veterinary professions strongly complement one another. The recent gaining acceptance among human and veterinary from humans (573), and domestic and exotic animals introduction of MALDI-TOF mass spectrometry is displaying great promise for the rapid, accurate and cost-effective diagnostic microbiology laboratories. (929) were analyzed by MS and included identification of bacterial and fungal pathogens. Integration of this proteomic signature-based approach into (human/animal isolates): staphylococci (112/193), contemporary laboratory practice holds great promise in revolutionizing clinical microbiology, as is evidenced by recent •The rapidity and accuracy of the MBT for human streptococci (61/112), enterococci (54/109), other applications in both human (Bizzini, 2012; Boggs, 2012; Carbonnelle, 2011; van Veen, 2012) and veterinary (Ahrholdt, (98.8%/94.4%, genus-/species-level, respectively) and Gram-positive cocci (36/1), Gram-positive bacilli 2011; Hijazin, 2011; Kuhnert, 2012; Vila, 2012) medicine. The improvement in laboratory turn-around time from 48-96 animal (95.4%/87.2%) pathogen identifications is (18/56), Enterobacteriaceae (147/252), Pseudomonas to 24 hours for pathogen identification can be expected to have real-time benefits in terms of earlier, targeted unsurpassed compared with traditional biochemical aeruginosa (61/69), other nonfermentative bacilli antimicrobial selection, detection of emerging diseases, and permitting of quarantine to be instituted sooner. methods. (56/49), Pasturella spp. (1/51), Actinobacillus spp. (0/9), and other Gram-negative bacilli (27/28). Isolates were In this study we evaluated the utility of MALDI-TOF mass spectrometry as a single platform for rapid pathogen •Infection and antimicrobial resistance health threats in identified by standard biochemical methods, and by MS identification in a regional USA laboratory practice routinely performing analyses on both human and animal-source humans and animals are often shared between these (Bruker Daltonics MALDI Biotyper™ v3.0) according to specimens. populations, and local and regional diagnostic the manufacturer’s recommendations and validation laboratories serve a critical public health sentinel algorithms. Table 1. Identification of 1,502 human- and animal-source isolates (year 2010) tested by MBT (Bruker Daltonics MALDI Biotyper™ v3.0). function in detecting their presence. RESULTS: Overall, MS provided species- and genus-level IDs, respectively, for 90.8% and 97.1% of 1,502 tested % Species Level ID % Genus Level ID % No ID •Utilization of MS for identification of pathogens recovered from human and animal specimens facilitates isolates. Among human and animal-source isolates, Isolate Groups (nspecies) nHuman nAnimal Human Animal Human Animal Human Animal respectively, MS identified 94.4%/87.2% to species- Coagulase-negative Staphylococcus (14) 52 48 88.5 62.5 9.6 37.5 1.9 0 and enhances the likelihood of detecting emerging level, 98.8%/95.4% to genus-level and 1.2%/4.6%, no Staphylococcus epidermidis 29 6 100 100 0 0 0 0 pathogens in both populations, and supports a key Staphylococcus felis 0 13 0 15.4 0 84.6 0 0 component of laboratory infrastructure essential for identification (not in database); compared with MS, Staphylococcus haemolyticus 2 2 100 0 0 100 0 0 promotion of the ‘One Medicine/One Health’ concept. standard ID approaches identified 70.0%/51.8% to Staphylococcus saprophyticus 7 0 71.4 0 28.6 0 0 0 species-level and 94.1%/88.2% to genus-level. Staphylococcus schleiferi 0 22 0 77.3 0 22.7 0 0 Discordant or missing IDs were present in 5.9% of Coagulase-positive Staphylococcus (4) 1 124 100 84.7 0 14.5 0 0.8 Staphylococcus intermedius group 1 124 100 84.7 0 14.5 0 0.8 human and 11.8% of animal source isolates. Staphylococcus aureus 59 21 100 100 0 0 0 0 Enterococcus species (8) 54 109 100 100 0 0 0 0 SELECTED REFERENCES CONCLUSION: Emerging infection health threats are Enterococcus faecalis 47 83 100 100 0 0 0 0 often zoonotic in origin, and local and regional Enterococcus faecium 6 19 100 100 0 0 0 0 Ahrholdt J, Murugaiyan J, Straubinger RK, Jagielski T, diagnostic laboratories serve a critical sentinel function Escherichia coli 49 104 100 100 0 0 0 0 Roesler U. Epidemiological analysis of worldwide bovine, Klebsiella species (3) 21 22 100 100 0 0 0 0 in detecting their presence. Utilization of MS for ID of canine and human clinical Prototheca isolates by PCR Klebsiella oxytoca 6 6 100 100 0 0 0 0 genotyping and MALDI-TOF mass spectrometry proteomic pathogens recovered from human and animal Klebsiella pneumoniae 14 16 100 100 0 0 0 0 specimens facilitates and enhances the likelihood of Proteus mirabilis 10 39 100 100 0 0 0 0 phenotyping. Med Mycol. 2011 Sep 12. [Epub ahead of detecting emerging pathogens in both populations. The Other Enterobacteriaceae (24) 67 87 95.5 92.0 4.5 4.6 0 3.4 print] accuracy and rapidity of MS technology for human Citrobacter species 13 13 92.3 76.9 7.7 7.7 0 15.4 Enterobacter species 15 12 93.3 100 6.7 0 0 0 (94.4%/98.8%, species-/genus-level) and animal (87.2%/ Bizzini A, Greub G. Matrix-assisted laser desorption Morganella morganii 6 7 100 100 0 0 0 0 ionization time-of-flight mass spectrometry, a revolution in 95.4%) pathogen IDs, supports a key component of Providencia species 3 5 100 100 0 0 0 0 laboratory infrastructure essential for promotion of the Salmonella species 10 16 100 93.8 0 6.2 0 0 clinical microbial identification. Clin Microbiol Infect. 2010 ‘One Medicine/One Health’ concept. Pseudomonas aeruginosa 61 69 100 100 0 0 0 0 Nov;16(11):1614-9. Other Non-fermentative GNB (38) 56 49 89.3 71.4 7.1 24.5 3.6 4.1 Achromobacter xylosoxidans 5 2 100 100 0 0 0 0 Boggs SR, Cazares LH, Drake RR. Characterisation of a *Revised to include additional isolates. Acinetobacter species 12 9 91.7 88.9 8.3 0 0 11.1 Other Pseudomonas species 10 14 80.0 57.1 0 42.9 20.0 0 Staphylococcus aureus USA300 protein signature using Stenotrophomonas maltophilia 17 6 100 83.3 0 16.7 0 0 matrix-assisted laser desorption/ionization time-of-flight Pasturella/Actinobacillus spp. (7) 1 60 100 78.3 0 8.3 0 13.3 mass spectrometry. J Med Microbiol. 2012 Feb 9. [Epub Actinobacillus species 0 11 0 72.7 0 9.1 0 18.2 ahead of print] Pasteurella canis 0 19 0 84.2 0 15.8 0 0 Pasteurella multocida 1 22 100 81.8 0 4.6 0 13.6 Gram-positive bacilli (27) 18 56 72.2 42.9 16.7 23.2 11.1 33.9 Carbonnelle E, Mesquita C, Bille E, Day N, Dauphin B, MATERIALS AND METHODS Arcanobacterium pyogenes 0 14 0 92.9 0 7.1 0 0 Beretti JL, Ferroni A, Gutmann L, Nassif X.
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