Abstract
Acanthamoeba spp. are ubiquitous in the environment and have the potential to cause severe infections. The different genotypes of Acanthamoeba have been shown to influence the severity of the disease and response to therapy. Characterizing Acanthamoeba spp. upto genotype can aid in infection control practices. Twenty-five Acanthamoeba isolates, characterized by 18S rDNA sequencing, were subjected to MALDI-TOF MS analysis by creating a database for the individual genotypes. The differentiating features of the various spectra were observed; the coded samples were then tested against the created database. The results of identification were compared with sequencing. Five different genotypes were obtained—T3, T4, T5, T10, and T11. Spectral analysis revealed genus-specific and genotype-specific peaks. The peak patterns for individual genotype were discrete and reproducible. Clinical isolates produced different peaks from the environmental isolate of the same genotype. A concordance of 92% was obtained with MALDI-TOF MS in comparison with 18sDNA sequencing. MALDI-TOF MS, once optimized, has the potential to reliably identify the genotype of Acanthamoeba spp. and to differentiate clinical isolate from mere contaminant.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arnalich-Montiel F, Lumbreras-fernández B, Martín-Navarro CM, Valladares B, Lopez-Velez R, Morcillo-Laiz R, Lorenzo-Morales J (2014) Influence of Acanthamoeba genotype on clinical course and outcomes for patients with Acanthamoeba keratitis in Spain. J Clin Microbiol 52(4):1213–1216. https://doi.org/10.1128/JCM.00031-14
Avila CC, Almeida FG, Palmisano G (2016) Direct identification of trypanosomatids by matrix-assisted laser desorption ionization—time of flight mass spectrometry (DIT MALDI-TOF MS). J Mass Spectrom 51(8):549–557. https://doi.org/10.1002/jms.3763
Brondfield M, Reid M, Rutishauser RL, Cope JR, Tang J, Ritter JM, Matanock A, Ali I, Doernberg SB, Hilts-Horeczko A, DeMarco T, Klein L, Babik JM (2017) Disseminated Acanthamoeba infection in a heart transplant recipient treated successfully with a miltefosine containing regimen: case report and review of the literature. Transpl Infect Dis 19(2):e12661. https://doi.org/10.1111/tid.12661.
Calderaro A, Piergianni M, Buttrini M, Montecchini S, Piccolo G, Gorrini C, Rossi S, Chezzi C, Arcangeletti MC, Medici MC, De Conto F (2015) MALDI-TOF mass spectrometry for the detection and differentiation of Entamoeba histolytica and Entamoeba dispar. PLoS One 10(4):e0122448. https://doi.org/10.1371/journal.pone.0122448
Chierico DF, Cave DD, Accardi C, Santoro M, Masotti A, D’Alfonso R, Berrilli F, Urbani A, Putignani L (2016) Identification and typing of free-living Acanthamoeba spp. by MALDI-TOF MS Biotyper. Exp Parasitol 170:82–89. https://doi.org/10.1016/j.exppara.2016.09.007
Clark AE, Kaleta EJ, Arora A, Wolk M (2013) Matrix-assisted laser desorption ionization—time of flight mass spectrometry: a fundamental shift in the routine practice of clinical microbiology. Clin Microbiol Rev 26(3):547–603. https://doi.org/10.1128/CMR.00072-12
Corsaro D, Venditti D (2011) More Acanthamoeba genotypes: limits to the use of rDNA fragments to describe new genotypes. Acta Protozool 50:49–54
da Rocha-Azevedo B, Costa e Silva-Filho F. (2007) Biological characterization of a clinical and an environmental isolate of Acanthamoeba polyphaga: analysis of relevant parameters to decode pathogenicity. Arch Microbiol 188(5):441–449. https://doi.org/10.1007/s00203-007-0264-3
Dykova I, Lom J, Schroeder-Diedrich JM, Booton GC, Byers TJ (1999) Acanthamoeba strains isolated from organs of freshwater fishes. J Parasitol 85(6):1106–1113. https://doi.org/10.2307/3285675
Fuerst PA, Booton GC, Crary M (2015) Phylogenetic analysis and the evolution of the 18S rRNA gene typing system of Acanthamoeba. J Eukaryot Microbiol 62(1):69–84. https://doi.org/10.1111/jeu.12186
Huang JM, Lin WC, Li SC, Shih MH, Chan WC, Shin JW, Huang FC (2016) Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain. Exp Parasitol 166:60–67. https://doi.org/10.1016/j.exppara.2016.03.018
Hugo ER, Stewart VJ, Gast RJ, Byers TJ (1992) Purification of amoeba mtDNA using the UNSET procedure. In: JJ Lee and AT Soldo (ed), Protocols in protozoology. p D-7.1–D-7.2.
Intelicato-Young J, Fox A (2013) Mass spectrometry and tandem mass spectrometry characterization of protein patterns, protein markers and whole proteomes for pathogenic bacteria. J Microbiol Methods 92(3):381–386. https://doi.org/10.1016/j.mimet.2013.01.004
Khan NA (2006) Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev 30(4):564–595. https://doi.org/10.1111/j.1574-6976.2006.00023.x
Lachaud L, Fernández-Arévalo A, Normand A et al (2017) Identification of Leishmania by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry using a free web-based application and a dedicated mass-spectral library. J Clin Microbiol 55(10):2924–2933. https://doi.org/10.1128/JCM.00845-17
Ledee DR, Booton GC, Awwad MH, Sharma S, Aggarwal RK, Niszl IA, Markus MB, Fuerst PA, Byers TJ (2003) Advantages of using mitochondrial 16S rDNA sequences to classify clinical isolates of Acanthamoeba. Investig Opthalmology Vis Sci 44(3):1142. https://doi.org/10.1167/iovs.02-0485
Liang SY, Ji DR, Hsia KT, Hung CC, Sheng WH, Hsu BM, Chen JS, MH W, Lai CH, Ji DD (2010) Isolation and identification of Acanthamoeba species related to amoebic encephalitis and nonpathogenic free-living amoeba species from the rice field. J Appl Microbiol 109(4):1422–1429. https://doi.org/10.1111/j.1365-2672.2010.04779.x
Maciver SK, Asif M, Simmen MW, Lorenzo-Morales J (2013) A systematic analysis of Acanthamoeba genotype frequency correlated with source and pathogenicity: T4 is confirmed as a pathogen-rich genotype. Eur J Protistol 49(2):217–221. https://doi.org/10.1016/j.ejop.2012.11.004
Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16(2):273–307. https://doi.org/10.1128/CMR.16.2.273
Moura H, Izquierdo F, Woolfitt AR, Wagner G, Pinto T, del Aguila C, Barr JR (2015) Detection of biomarkers of pathogenic Naegleria fowleri through mass spectrometry and proteomics. J Eukaryot Microbiol 62(1):12–20. https://doi.org/10.1111/jeu.12178
Schroeder JM, Booton GC, Hay J, Niszl IA, Seal DV, Markus MB, Fuerst PA, Byers TJ (2001) Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. J Clin Microbiol 39(5):1903–1911. https://doi.org/10.1128/JCM.39.5.1903-1911.2001
Sharma M, Sharma S, Sharma S, Megha K, Nada R, Khurana S (2017) Osteo-cutaneous acanthamoebiasis in a non-immunocompromised patient with a favorable outcome. Parasitol Int 66(6):727–730. https://doi.org/10.1016/j.parint.2017.08.003
Stothard D, Schroeder-Diedrich J, Awwad MH, Gast RJ, Ledee DR, Rodriguez-Zaragoza S, Dean CL, Fuerst PA, Byers TJ (1998) The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45(1):45–54. https://doi.org/10.1111/j.1550-7408.1998.tb05068.x
Tawfeek GM, Bishara SA, Sarhan RM, ElShabrawi TE, ElSaady Khayyal A (2016) Genotypic, physiological, and biochemical characterization of potentially pathogenic Acanthamoeba isolated from the environment in Cairo, Egypt. Parasitol Res 115(5):1871–1881. https://doi.org/10.1007/s00436-016-4927-3
Tu EY, Joslin CE, Sugar J, Shoff ME, Booton GC (2008) Prognostic factors affecting visual outcome in Acanthamoeba keratitis. Ophthalmology 115(11):1998–2003. https://doi.org/10.1016/j.ophtha.2008.04.038.Prognostic
Yssouf A, Almeras L, Raoult D, Parola P (2016) Emerging tools for identification of arthropod vectors. Future Microbiol 11(4):549–566. https://doi.org/10.2217/fmb.16.5
Funding
The funding for this work was received from Indian Council of Medical Research vide No. 5/3/3/33/2013-ECD-I.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Megha, K., Sharma, M., Gupta, A. et al. Protein profiling of Acanthamoeba species using MALDI-TOF MS for specific identification of Acanthamoeba genotype. Parasitol Res 117, 729–736 (2018). https://doi.org/10.1007/s00436-017-5743-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-017-5743-0