Analysis of Cryptosporidium spp. from clinical samples by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry.

Cryptosporidium MALDI-TOF mass spectrometry matrix-assisted laser desorption/ionization time-of-flight purification

Journal

Journal of applied microbiology
ISSN: 1365-2672
Titre abrégé: J Appl Microbiol
Pays: England
ID NLM: 9706280

Informations de publication

Date de publication:
Oct 2021
Historique:
revised: 09 03 2021
received: 19 01 2021
accepted: 13 03 2021
pubmed: 19 3 2021
medline: 16 10 2021
entrez: 18 3 2021
Statut: ppublish

Résumé

To purify Cryptosporidium spp. oocysts from clinical stool samples and evaluate using an up-to-date mass spectrometry protocol producing high-quality reference spectra. A refined purification protocol was developed for oocysts from stools, involving salt flotation and potassium bromide density centrifugation. Purified oocysts were prepared for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) by formic acid extraction, and the extracts analysed using the Bruker MALDI Biotyper system. Individual spectral markers were identified by their specific mass peaks. Cryptosporidium parvum oocysts (Iowa strain) propagated in vivo, and C.  parvum (n = 2) and Cryptosporidium hominis (n = 1) oocysts from clinical stool samples produced distinct spectra that were considered specific to Cryptosporidium spp. with no evidence of contamination. The production of distinct spectra demonstrated the utility of the purification method for oocysts from clinical stool samples and provided reference spectra. The use of MALDI-TOF MS and other mass spectrometry techniques has been limited previously to C.  parvum oocysts propagated in vivo. Appropriate purification of oocysts can achieve sufficient biomass, enabling analysis by MALDI-TOF MS and potentially other mass spectrometry platforms, facilitating peptide and protein discovery and identification of biomarkers from a much wider range of Cryptosporidium spp. from natural infections.

Identifiants

pubmed: 33735496
doi: 10.1111/jam.15077
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

1840-1847

Subventions

Organisme : Society for Applied Microbiology

Informations de copyright

© 2021 The Society for Applied Microbiology.

Références

Alberts, B., Johnson, A., Lewis, J. and Al, E. (2002) Analyzing protein structure and function. New York: Garland Science.
Aldred, S., Grant, M.M. and Griffiths, H.R. (2004) The use of proteomics for the assessment of clinical samples in research. Clin Biochem 37, 943-952.
Alves, M., Xiao, L., Sulaiman, I., Lal, A.A., Matos, O. and Antunes, F. (2003) Subgenotype analysis of Cryptosporidium isolates from humans, cattle, and zoo ruminants in Portugal. J Clin Microbiol 41, 2744-2747.
Barrett, J., Jefferies, J.R. and Brophy, P.M. (2000) Parasite proteomics. Parasitol Today 16, 400-403.
Bhalchandra, S., Cardenas, D. and Ward, H.D. (2018) Recent breakthroughs and ongoing limitations in Cryptosporidium research. F1000Research, 7, F1000 Faculty Rev-1380
Cama, V.A., Bern, C., Roberts, J., Cabrera, L., Sterling, C.R., Ortega, Y., Gilman, R.H. and Xiao, L. (2008) Cryptosporidium species and subtypes and clinical manifestations in children, Peru. Emerg Infect Dis 14, 1567-1574.
Chalmers, R.M., Elwin, K., Thomas, A.L., Guy, E.C. and Mason, B. (2009) Long-term Cryptosporidium typing reveals the aetiology and species-specific epidemiology of human cryptosporidiosis in England and Wales, 2000 to 2003. Euro Surveill 14, 19086.
Checkley, W., White, A.C. Jr, Jaganath, D., Arrowood, M.J., Chalmers, R.M., Chen, X.M., Fayer, R., Griffiths, J.K. et al. (2015) A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for cryptosporidium. Lancet Infect Dis 15, 85-94.
Crutchfield, C.A., Thomas, S.N., Sokoll, L.J. and Chan, D.W. (2016) Advances in mass spectrometry-based clinical biomarker discovery. Clin Proteomics 13, 1.
Entrala, E., Molina-Molina, J., Rosales-Lombardo, M., Sanchez-Moreno, M. and Mascaro-Lazcano, C. (2000) Cryptosporidium parvum: oocysts purification using potassium bromide discontinuous gradient. Vet Parasitol 92, 223-226.
Glassmeyer, S.T., Ware, M.W., Schaefer, F.W. 3rd, Shoemaker, J.A. and Kryak, D.D. (2007) An improved method for the analysis of Cryptosporidium parvum oocysts by matrix-assisted laser desorption/ionization time of flight mass spectrometry. J Eukaryot Microbiol 54, 479-481.
Haserick, J.R., Leon, D.R., Samuelson, J. and Costello, C.E. (2017) Asparagine-linked glycans of Cryptosporidium parvum contain a single long arm, are barely processed in the endoplasmic reticulum (ER) or golgi, and show a strong bias for sites with threonine. Mol Cell Proteomics 16, s42-s53.
Haslam, C., Hellicar, J., Dunn, A., Fuetterer, A., Hardy, N., Marshall, P., Paape, R., Pemberton, M. et al. (2016) The evolution of MALDI-TOF mass spectrometry toward ultra-high-throughput screening: 1536-well format and beyond. J Biomol Screen 21, 176-186.
Jarnuczak, A.F., Lee, D.C., Lawless, C., Holman, S.W., Eyers, C.E. and Hubbard, S.J. (2016) Analysis of Intrinsic Peptide Detectability via Integrated Label-Free and SRM-Based Absolute Quantitative Proteomics. J Proteome Res 15, 2945-2959.
Jean Beltran, P.M., Federspiel, J.D., Sheng, X. and Cristea, I.M. (2017) Proteomics and integrative omic approaches for understanding host-pathogen interactions and infectious diseases. Mol Syst Biol 13, 922.
Khalil, I.A., Troeger, C., Rao, P.C., Blacker, B.F., Brown, A., Brewer, T.G., Colombara, D.V., De Hostos, E.L. et al. (2018) Morbidity, mortality, and long-term consequences associated with diarrhoea from Cryptosporidium infection in children younger than 5 years: a meta-analyses study. Lancet Glob Health 6, e758-e768.
Lee, S.U., Joung, M., Nam, T., Park, W.Y., Ji, Y.H. and Yu, J.R. (2011) Cryptosporidium parvum: radiation-induced alteration of the oocyst proteome. Exp Parasitol 127, 25-30.
Magnuson, M.L., Owens, J.H. and Kelty, C.A. (2000) Characterization of Cryptosporidium parvum by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Appl Environ Microbiol 66, 4720-4724.
Manque, P.A., Tenjo, F., Woehlbier, U., Lara, A.M., Serrano, M.G., Xu, P., Alves, J.M., Smeltz, R.B. et al. (2011) Identification and immunological characterization of three potential vaccinogens against Cryptosporidium species. Clin Vaccine Immunol 18, 1796-1802.
Matsubayashi, M., Kimata, I., Iseki, M., Lillehoj, H.S., Matsuda, H., Nakanishi, T., Tani, H., Sasai, K. et al. (2005) Cross-reactivities with Cryptosporidium spp. by chicken monoclonal antibodies that recognize avian Eimeria spp. Vet Parasitol 128, 47-57.
Matsubayashi, M., Teramoto-Kimata, I., Uni, S., Lillehoj, H.S., Matsuda, H., Furuya, M., Tani, H. and Sasai, K. (2013) Elongation factor-1 alpha is a novel protein associated with host cell invasion and a potential protective antigen of Cryptosporidium parvum. J Biol Chem 288, 34111-34120.
Robinson, G., Elwin, K. and Chalmers, R.M. (2020) Cryptosporidium diagnostic assays: molecular detection. Methods Mol Biol 2052, 11-22.
Ryan, U., Zahedi, A. and Paparini, A. (2016) Cryptosporidium in humans and animals - a one health approach to prophylaxis. Parasite Immunol 38, 535-547.
Ryley, J.F., Meade, R., Hazelhurst, J. and Robinson, T.E. (1976) Methods in coccidiosis research: separation of oocysts from faeces. Parasitology 73, 311-326.
Sanderson, S.J., Xia, D., Prieto, H., Yates, J., Heiges, M., Kissinger, J.C., Bromley, E., Lal, K. et al. (2008) Determining the protein repertoire of Cryptosporidium parvum sporozoites. Proteomics 8, 1398-1414.
Siddiki, A.Z. (2013) Sporozoite proteome analysis of Cryptosporidium parvum by one-dimensional SDS-PAGE and liquid chromatography tandem mass spectrometry. J Vet Sci 14, 107-114.
Tandel, J., English, E.D., Sateriale, A., Gullicksrud, J.A., Beiting, D.P., Sullivan, M.C., Pinkston, B. and Striepen, B. (2019) Life cycle progression and sexual development of the apicomplexan parasite Cryptosporidium parvum. Nat Microbiol 4, 2226-2236.
Truong, Q. and Ferrari, B.C. (2006) Quantitative and qualitative comparisons of Cryptosporidium faecal purification procedures for the isolation of oocysts suitable for proteomic analysis. Int J Parasitol 36, 811-819.
Webster, J. and Oxley, D. (2012) Protein identification by MALDI-TOF mass spectrometry. Methods Mol Biol 800, 227-240.

Auteurs

R Gathercole (R)

School of Biomedical, Nutritional and Sport Sciences, School of Biomedical, Nutritional and Sport Sciences, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.

E Tranfield (E)

Bruker Microbiology & Diagnostics, Bruker, Coventry, UK.

D Xia (D)

Comparative Biomedical Sciences, Royal Veterinary College, University of London, London, UK.

G Perez-Cordon (G)

Cryptosporidium Reference Unit, Public Health Wales Microbiology and Health Protection, Singleton Hospital, Swansea, Wales, UK.
Swansea University Medical School, Singleton Park, Swansea, Wales, UK.

G Robinson (G)

Cryptosporidium Reference Unit, Public Health Wales Microbiology and Health Protection, Singleton Hospital, Swansea, Wales, UK.
Swansea University Medical School, Singleton Park, Swansea, Wales, UK.

D Timofte (D)

Institute of Infection, Veterinary and Ecological Sciences, Department of Veterinary Anatomy, Physiology and Pathology, University of Liverpool, Cheshire, UK.

F Zendri (F)

Institute of Infection, Veterinary and Ecological Sciences, Department of Veterinary Anatomy, Physiology and Pathology, University of Liverpool, Cheshire, UK.

R M Chalmers (RM)

Cryptosporidium Reference Unit, Public Health Wales Microbiology and Health Protection, Singleton Hospital, Swansea, Wales, UK.
Swansea University Medical School, Singleton Park, Swansea, Wales, UK.

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