BMC Research Notes | |
Leptospira species and serovars identified by MALDI-TOF mass spectrometry after database implementation | |
Maria Cristina Medici1  Carlo Chezzi1  Maria Cristina Arcangeletti1  Flora De Conto1  Maddalena Piergianni1  Sabina Rossi1  Mirko Buttrini1  Sara Montecchini1  Chiara Gorrini1  Giovanna Piccolo1  Adriana Calderaro1  | |
[1] Unit of Microbiology and Virology, Department of Clinical and Experimental Medicine, University of Parma, Viale A. Gramsci, 14-43126 Parma, Italy | |
关键词: Database implementation; Identification; MALDI-TOF MS; Leptospira sp.; | |
Others : 1132651 DOI : 10.1186/1756-0500-7-330 |
|
received in 2014-05-17, accepted in 2014-05-21, 发布年份 2014 | |
【 摘 要 】
Background
Leptospirosis, a spirochaetal zoonotic disease of worldwide distribution, endemic in Europe, has been recognized as an important emerging infectious disease, though yet it is mostly a neglected disease which imparts its greatest burden on impoverished populations from developing countries. Leptospirosis is caused by the infection with any of the more than 230 serovars of pathogenic Leptospira sp. In this study we aimed to implement the MALDI-TOF mass spectrometry (MS) database currently available in our laboratory with Leptospira reference pathogenic (L. interrogans, L. borgpetersenii, L. kirschneri, L. noguchii), intermediate (L. fainei) and saprophytic (L. biflexa) strains of our collection in order to evaluate its possible application to the diagnosis of leptospirosis and to the typing of strains. This was done with the goal of understanding whether this methodology could be used as a tool for the identification of Leptospira strains, not only at species level for diagnostic purposes, but also at serovar level for epidemiological purposes, overcoming the limits of serological and molecular conventional methods. Twenty Leptospira reference strains were analysed by MALDI-TOF MS. Statistical analysis of the protein spectra was performed by ClinProTools software.
Results
The spectra obtained by the analysis of the reference strains tested were grouped into 6 main classes corresponding to the species analysed, highlighting species-specific protein profiles. Moreover, the statistical analysis of the spectra identified discriminatory peaks to recognize Leptospira strains also at serovar level extending previously published data.
Conclusions
In conclusion, we confirmed that MALDI-TOF MS could be a powerful tool for research and diagnostic in the field of leptospirosis with broad applications ranging from the detection and identification of pathogenic leptospires for diagnostic purposes to the typing of pathogenic and non-pathogenic leptospires for epidemiological purposes in order to enrich our knowledge about the epidemiology of the infection in different areas and generate control strategies.
【 授权许可】
2014 Calderaro et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20150304033743693.pdf | 1228KB | download | |
Figure 3. | 70KB | Image | download |
Figure 2. | 64KB | Image | download |
Figure 1. | 90KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
【 参考文献 】
- [1]Bharti AR, Nally JE, Ricaldi JN, Matthias MA, Diaz MM, Lovett MA, Levett PN, Gilman RH, Willig MR, Gotuzzo E, Vinetz JM, Peru-United States Leptospirosis Consortium: Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 2003, 3:757-771.
- [2]Ko AI, Goarant C, Picardeau M: Leptospira: the dawn of the molecular genetics era for an emerging zoonotic pathogen. Nat Rev Microbiol 2009, 7:736-747.
- [3]Levett PN: Leptospirosis. Clin Microbiol Rev 2001, 4:296-326.
- [4]Adler B, Lo M, Seemann T, Murray GL: Pathogenesis of leptospiroses: the influence of genomics. Vet Microbiol 2011, 153:73-81.
- [5]Brenner D, Kaufmann AF, Sulzer KR, Steigerwalt AG, Rogers FC, Weyant RS: Further determination of DNA relatedness between serogroups and serovars in the family Leptospiraceae with a proposal for Leptospira alexanderi sp. nov. and four new Leptospira genomospecies. Int J Syst Bacteriol 1999, 49:839-858.
- [6]Cerqueira GM, Picardeau M: A century of Leptospira strain typing. Infect Genet Evol 2009, 9:760-768.
- [7]Djelouadji Z, Roux V, Raoult D, Kodjo A, Drancourt M: Rapid MALDI-TOF mass spectrometry identification of Leptospira organisms. Vet Microbiol 2012, 158:142-146.
- [8]Postic D, Merien F, Perolat P, Baranton G: Laboratory methods - Biological diagnosis for Leptospirosis. In Biological diagnosis Leptospirosis - Lyme borreliosis. 2nd enlarged edition. Paris: Institut Pasteur; 2000:139-196.
- [9]Perolat P, Chappel RJ, Adler B, Baranton G, Bulach DM, Billinghurst ML, Letocart M, Merien F, Serrano MS: Leptospira fainei sp. nov., isolated from pigs in Australia. Int J Syst Bacteriol 1998, 48:851-858.
- [10]Ciceroni L, Ciarrocchi S, Ciervo A, Petrucca A, Pinto A, Calderaro A, Viani I, Galati L, Dettori G, Chezzi C: Differentiation of leptospires of the serogroup Pomona by monoclonal antibodies, pulsed-field gel electrophoresis and arbitrarily primed polymerase chain reaction. Res Microbiol 2002, 153:37-44.
- [11]Herrmann JL, Bellenger E, Perolat P, Baranton G, Saint Girons I: Pulsed-field gel electrophoresis of NotI digests of leptospiral DNA: a new rapid method of serovar identification. J Clin Microbiol 1992, 30:1696-1702.
- [12]Zuerner RL, Ellis WA, Bolin CA, Montgomery JM: Restriction fragment length polymorphisms distinguish Leptospira borgpetersenii serovar hardjo type hardjo-bovis isolates from different geographical locations. J Clin Microbiol 1993, 31:578-583.
- [13]Majed Z, Bellenger E, Postic D, Pourcel C, Baranton G, Picardeau M: Identification of variable-number tandem-repeat loci in Leptospira interrogans sensu stricto. J Clin Microbiol 2005, 43:539-545.
- [14]Ahmed N, Devi SM, Valverde M, Vijayachari P, Machang’u RS, Ellis WA, Hartskeerl RA: Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species. Ann Clin Microbiol Antimicrob 2006, 5:28. BioMed Central Full Text
- [15]Branger C, Blanchard B, Fillonneau C, Suard I, Aviat F, Chevallier B, André-Fontaine G: Polymerase chain reaction assay specific for pathogenic Leptospira based on the gene hap1 encoding the hemolysis-associated protein-1. FEMS Microbiol Lett 2005, 243:437-445.
- [16]La Scola B, Bui LT, Baranton G, Khamis A, Raoult D: Partial rpoB gene sequencing for identification of Leptospira species. FEMS Microbiol Lett 2006, 263:142-147.
- [17]Merien F, Amouriaux P, Perolat P, Baranton G, Saint Girons I: Polymerase chain reaction for detection of Leptospira sp. in clinical samples. J Clin Microbiol 1992, 30:2219-2224.
- [18]Natarajaseenivasan K, Vijayachari P, Sharma S, Sugunan AP, Vedhagiri K, Selvin J, Sehgal SC: FlaB PCR-based identification of pathogenic leptospiral isolates. J Microbiol Immunol Infect 2010, 43:62-69.
- [19]Slack AT, Symonds ML, Dohnt MF, Smythe LD: Identification of pathogenic Leptospira species by conventional or real-time PCR and sequencing of the DNA gyrase subunit B encoding gene. BMC Microbiol 2006, 6:95. BioMed Central Full Text
- [20]El-Khéchine A, Couderc C, Flaudrops C, Raoult D, Drancourt M: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification of mycobacteria in routine clinical practice. PLoS One 2011, 6:e24720.
- [21]Seng P, Drancourt M, Gouriet F, La Scola B, Fournier PE, Rolain JM, Raoult D: Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis 2009, 15:543-551.
- [22]Calderaro A, Piccolo G, Montecchini S, Buttrini M, Gorrini C, Rossi S, Arcangeletti MC, De Conto F, Medici MC, Chezzi C: MALDI-TOF MS analysis of human and animal Brachyspira species and benefits of database extension. J Prot 2013, 78:273-280.
- [23]Calderaro A, Gorrini C, Piccolo G, Montecchini S, Buttrini M, Rossi S, Piergianni M, Arcangeletti MC, De Conto F, Chezzi C, Medici MC: Identification of Borrelia species after creation of an in-house MALDI-TOF MS database. PLoS One 2014, 12(2):e88895.
- [24]Rettinger A, Krupka I, Grünwald K, Dyachenko V, Fingerle V, Konrad R, Raschel H, Busch U, Sing A, Straubinger RK, Huber I: Leptospira sp. strain identification by MALDI TOF MS is an equivalent tool to 16S rRNA gene sequencing and multi locus sequence typing (MLST). BMC Microbiol 2012, 12:185. BioMed Central Full Text
- [25]Ayyadurai S, Flaudrops C, Raoult D, Drancourt M: Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDITOF) mass spectrometry. BMC Microbiol 2010, 10:285. BioMed Central Full Text