Background Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected tropical poverty-linked illness. This infection is endemic of Andean regions and it is estimated that approximately 1.7 million of South Americans are at risk. This bacterium is a fastidious slow growing microorganism, which is difficult and cumbersome to isolate from clinical sources, thereby hindering the availability of phylogenetic relationship of clinical samples. The aim of this study was to perform Multi Locus Sequence Typing of B. bacilliformis directly in blood from patients diagnosed with Oroya fever during an outbreak in Northern Peru. Methodology/Principal Findings DNA extracted among blood samples from patients diagnosed with Oroya’s fever were analyzed with MLST, with the amplification of 7 genetic loci (ftsZ, flaA, ribC, rnpB, rpoB, bvrR and groEL) and a phylogenetic analysis of the different Sequence Types (ST) was performed. A total of 4 different ST were identified. The most frequently found was ST1 present in 66% of samples. Additionally, two samples presented a new allelic profile, belonging to new STs (ST 9 and ST 10), which were closely related to ST1. Conclusions/Significance The present data demonstrate that B. bacilliformis MLST studies may be possible directly from blood samples, being a promising approach for epidemiological studies. During the outbreak the STs of B. bacilliformis were found to be heterogeneous, albeit closely related, probably reflecting the evolution from a common ancestor colonizing the area. Additional studies including new samples and areas are needed, in order to obtain better knowledge of phylogenetic scenario B. bacilliformis.
References
[1]
Ihler GM. Bartonella bacilliformis: dangerous pathogen slowly emerging from deep background. FEMS Microbiol Lett 1996; 144:1–11. pmid:8870245 doi: 10.1016/0378-1097(96)00307-2
[2]
Sanchez Clemente N, Ugarte-Gil CA, Magui?a C, Pachas P, Blazes D, Bailey R et al. Bartonella bacilliformis: a systematic review of the literature to guide the research agenda for elimination. PLoS Negl Trop Dis. 2012; 6:e1819. doi: 10.1371/journal.pntd.0001819. pmid:23145188
[3]
Pachas PE. Enfermedad de Carrión (Bartonellosis) en el Peru. Lima. Ministerio de Salud, OGE, INS. 2001.
[4]
Magui?a C, Guerra H, Ventosilla P. Bartonellosis. Clin Dermatol. 2009; 27:271–280. doi: 10.1016/j.clindermatol.2008.10.006. pmid:19362689
[5]
Ticona E, Huaroto L, Garcia Y, Vargas L, Madariaga MG The pathophysiology of the acute phase of human bartonellosis resembles AIDS. Med Hypotheses 2010; 74:45–49. doi: 10.1016/j.mehy.2009.06.054. pmid:19665314
[6]
Magui?a C, Gotuzzo E Bartonellosis—new and old. Infect Dis Clin North Am.2000; 14:1–22. pmid:10738670 doi: 10.1016/s0891-5520(05)70215-4
[7]
Chamberlin J, Laughlin LW, Romero S, Solorzano N, Gordon S, Andre RG et al. Epidemiology of endemic Bartonella bacilliformis: A prospective cohort study in a Peruvian mountain valley community. J Infect Dis 2002; 186: 983–990. pmid:12232839 doi: 10.1086/344054
[8]
Birtles RJ, Fry NK, Ventosilla P, Cáceres AG, Sánchez E, Vizcarra H et al. Identification of Bartonella bacilliformis genotypes and their relevance to epidemiological investigations of human bartonellosis. J Clin Microbiol. 2002; 40: 3606–3612. pmid:12354853 doi: 10.1128/jcm.40.10.3606-3612.2002
[9]
Hambuch TM, Handley SA, Ellis B, Chamberlin J, Romero S, Regnery R. Population genetic analysis of Bartonella bacilliformis isolates from areas of Peru where Carrion's disease is endemic and epidemic. J Clin Microbiol. 2004; 42:3675–3680. pmid:15297516 doi: 10.1128/jcm.42.8.3675-3680.2004
[10]
Padilla RC, Ventura EG Genotipificación de aislamientos de Bartonella bacilliformis por amplificación de elementos repetitivos mediante el uso de REP-PCR y ERIC-PCR. Rev Peru Med Exp Clin 2003; 20: 128–131.
[11]
Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, Urwin R, Zhang Q et al. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci USA. 1998; 95: 3140–3145. pmid:9501229 doi: 10.1073/pnas.95.6.3140
[12]
Iredell J, Blanckenberg D, Arvand M, Grauling S, Feil EJ, Birtles RJ. Characterization of the natural population of Bartonella henselae by multilocus sequence typing. J Clin Microbiol. 2003; 41: 5071–5079. pmid:14605141 doi: 10.1128/jcm.41.11.5071-5079.2003
[13]
Arvand M, Raoult D, Feil EJ. Multi-locus sequence typing of a geographically and temporally diverse sample of the highly clonal human pathogen Bartonella quintana. PLoS One 2010; 5: e9765. doi: 10.1371/journal.pone.0009765. pmid:20333257
[14]
Bai Y, Malania L, Alvarez Castillo D, Moran D, Boonmar S, Chanlun A et al. Global distribution of Bartonella infections in domestic bovine and characterization of Bartonella bovis strains using multi-locus sequence typing. PLoS One 2013; 21;8:e80894. doi: 10.1371/journal.pone.0080894. pmid:24278342
[15]
Mullins KE, Hang J, Jiang J, Leguia M, Kasper MR, Magui?a C et al. Molecular typing of "Candidatus Bartonella ancashi," a new human pathogen causing verruga peruana. J Clin Microbiol 2013; 51:3865–3868 doi: 10.1128/JCM.01226-13. pmid:23985925
[16]
Chaloner GL, Ventosilla Palmira, Birtles RJ. Multi-Locus sequence analysis reveals profound genetic diversity among isolates of the human pathogen Bartonella bacilliformis. PLoS Negl Trop Dis. 2011; 5: e1248. doi: 10.1371/journal.pntd.0001248. pmid:21811647
[17]
del Valle Mendoza J, Silva Caso W, Tinco Valdez C, Pons MJ, del Valle LJ, Casabona Oré V et al. (2014) Diagnosis of Carrion's disease by direct blood PCR in thin blood smear negative samples. PLoS One 20;9:e92283. doi: 10.1371/journal.pone.0092283. pmid:24651298
[18]
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011; 28: 2731–9. doi: 10.1093/molbev/msr121. pmid:21546353
[19]
Chan KS, Kosoy M Analysis of multi-strain Bartonella pathogens in natural host population—do they behave as species or minor genetic variants? Epidemics 2010; 2: 165–172. doi: 10.1016/j.epidem.2010.08.002. pmid:21352787
[20]
Gomes C, Silva W, Tinco C, Martínez-Puchol , Pons MJ et al. Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability. Int J Infect Dis 2014; 21: 367. doi: 10.1016/j.ijid.2014.03.1177
[21]
Magui?a C, Gotuzzo E, Carcelén A, Salinas C, Cok J, Recavarren S et al. Compromiso gastrointestinal en bartonelosis o enfermedad de Carrión. Rev Gastroent Perú 1997; 17:31–43.
[22]
Pachas P. Epidemiología de la bartonelosis en la Región Chavín. Libro de resúmenes de trabajos libres. IX Congreso Nacional de Medicina Interna. Lima 1996.
[23]
Tarazona A, Magui?a C, López de Guimaraes D, Montoya M, Pachas PTerapia antibiótica para el manejo de la bartonelosis o enfermedad de Carrión en el Perú. Rev Peru Med Exp Salud Publ. 2006; 23:188–200.
[24]
Silva-Caso W, Pons MJ, Ruiz J, del Valle-Mendoza J. Antibiotic resistance in Bartonella bacilliformis clinical isolates from endemic area of Peru. J Global Antimicrob Resist 2015; 3: 222–223. doi: 10.1016/j.jgar.2015.05.004