1887

Abstract

Pythiosis is an infection of humans and other animals caused by the fungal-like pathogen . This pathogen causes life-threatening infection in the infected hosts. Culture, histopathology, serology and molecular tools are used to diagnose its infections. Successful management of pythiosis is directly linked to an early diagnosis. Thus, a rapid identification of putative cultures developing submerged sparsely septate hyphae is of extreme importance. However, few laboratories are familiar with the culture identification of this unique pathogen and its differential diagnosis with similar filamentous fungi.

We have evaluated the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) on 53 isolates of collected from cases of human and animal pythiosis in the USA and around the world. To assess the specificity of the approach, 18 pathogenic and saprotrophic filamentous fungal and fungal-like microbes were also tested.

MALDI-TOF in-house spectra correctly identified the 53 . isolates (score range 1.93–2.51). MALDI-TOF based identification within isolates showed protein spectra variation between geographical diverse isolates. A mass spectrometry approach was able to discriminate from the 18 filamentous fungal and fungal-like microbes in this study, including four spp. and plant pathogenic species.

The data showed MALDI-TOF could be used for the accurate and rapid culture identification of in the clinical laboratory.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000941
2019-04-01
2019-10-15
Loading full text...

Full text loading...

References

  1. Gaastra W, Lipman LJ, De Cock AW, Exel TK, Pegge RB et al. Pythium insidiosum: an overview. Vet Microbiol 2010;146:1–16 [CrossRef]
    [Google Scholar]
  2. Supabandhu J, Fisher MC, Mendoza L, Vanittanakom N. Isolation and identification of the human pathogen Pythium insidiosum from environmental samples collected in Thai agricultural areas. Med Mycol 2008;46:41–52 [CrossRef]
    [Google Scholar]
  3. Adl SM, Simpson AG, Lane CE, Lukeš J, Bass D et al. The revised classification of eukaryotes. J Eukaryot Microbiol 2012;59:429–514 [CrossRef]
    [Google Scholar]
  4. Chitasombat MN, Petchkum P, Horsirimanont S, Sornmayura P, Chindamporn A et al. Vascular pythiosis of carotid artery with meningitis and cerebral septic emboli: a case report and literature review. Med Mycol Case Rep 2018;21:57–62 [CrossRef]
    [Google Scholar]
  5. Thianprasit M, Chaiprasert A, Imwidthaya P. Human pythiosis. Curr Top Med Mycol 1996;7:43–54
    [Google Scholar]
  6. Mendoza L, Kaufman L, Mandy W, Glass R. Serodiagnosis of human and animal pythiosis using an enzyme-linked immunosorbent assay. Clin Diag Lab Immunol 1997;4:715–718
    [Google Scholar]
  7. Mendoza L, Nicholson V, Prescott JF. Immunoblot analysis of the humoral immune response to Pythium insidiosum in horses with pythiosis. J Clin Microbiol 1992;30:2980–2983
    [Google Scholar]
  8. Krajaejun T, Kunakorn M, Niemhom S, Chongtrakool P, Pracharktam R. Development and evaluation of an in-house enzyme-linked immunosorbent assay for early diagnosis and monitoring of human pythiosis. Clin Diagn Lab Immunol 2002;9:378–382 [CrossRef]
    [Google Scholar]
  9. Brown CC, McClure JJ, Triche P, Crowder C. Use of immunohistochemical methods for diagnosis of equine pythiosis. Am J Vet Res 1988;49:1866–1868
    [Google Scholar]
  10. Grooters AM, Gee MK. Development of a nested polymerase chain reaction assay for the detection and identification of Pythium insidiosum. J Vet Intern Med 2002;16:147–152 [CrossRef]
    [Google Scholar]
  11. Schurko AM, Mendoza L, de Cock AW, Bedard JE, Klassen GR. Development of a species-specific probe for Pythium insidiosum and the diagnosis of pythiosis. J Clin Microbiol 2004;42:2411–2418 [CrossRef]
    [Google Scholar]
  12. Badenoch PR, Coster DJ, Wetherall BL, Brettig HT, Rozenbilds MA et al. Pythium insidiosum keratitis confirmed by DNA sequence analysis. Br J Ophthalmol 2001;85:502–503 [CrossRef]
    [Google Scholar]
  13. Keeratijarut A, Lohnoo T, Yingyong W, Rujirawat T, Srichunrusami C et al. Detection of the oomycete Pythium insidiosum by real-time PCR targeting the gene coding for exo-1,3-β-glucanase. J Med Microbiol 2015;64:971–977 [CrossRef]
    [Google Scholar]
  14. Vilela R, Viswanathan P, Mendoza LA. A biochemical screening approach to putatively differentiate mammalian pathogenic Oomycota species in the clinical laboratory. J Med Microbiol 2015;64:862–868 [CrossRef]
    [Google Scholar]
  15. Moraes-Tondolo JS, Loreto ES, Denardi LB, Nunes-Mario DA, Alves SH et al. A simple, rapid and inexpensive screening method for the identification of Pythium insidiosum. J Microbiol Methods 2013;93:52–54 [CrossRef]
    [Google Scholar]
  16. Erhard M, Hipler UC, Burmester A, Brakhage AA, Wöstemeyer J. Identification of dermatophyte species causing onychomycosis and tinea pedis by MALDI-TOF mass spectrometry. Exp Dermatol 2008;17:356–361 [CrossRef]
    [Google Scholar]
  17. Hettick JM, Green BJ, Buskirk AD, Kashon ML, Slaven JE et al. Discrimination of Aspergillus isolates at the species and strain level by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry fingerprinting. Analyt Biochem 2008;380:276–281 [CrossRef]
    [Google Scholar]
  18. Alanio A, Beretti JL, Dauphin B, Mellado E, Quesne G et al. Matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry for fast and accurate identification of clinically relevant Aspergillus species. Clin Microbiol Infec 2011;17:750–755 [CrossRef]
    [Google Scholar]
  19. Krajaejun T, Lohnoo T, Jittorntam P, Srimongkol A, Kumsang Y et al. Assessment of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification and biotyping of the pathogenic oomycete Pythium insidiosum. Int J Infect Dis 2018;77:61–67 [CrossRef]
    [Google Scholar]
  20. Alejandro Rojas J, Jacobs JL, Napieralski S, Karaj B, Bradley CA et al. Oomycete species associated with soybean seedlings in North America—Part I: identification and pathogenicity characterization. Phytopathol 2017;107:280–292 [CrossRef]
    [Google Scholar]
  21. Lau AF, Drake SK, Calhoun LB, Henderson CM, Zelazny AM. Development of a clinically comprehensive database and a simple procedure for identification of molds from solid media by matrix-assisted laser desorption Ionization–Time of flight mass spectrometry. J Clin Microbiol 2013;51:828–834 [CrossRef]
    [Google Scholar]
  22. Bruker Daltonics Custom MSP and library creation. Revision C Billerica, MA: Bruker Daltonics; 2014
  23. Alizadeh M, Kolecka A, Boekhout T, Zarrinfar H, Ghanbari-Nahzag MA et al. Identification of Candida species isolated from vulvovaginitis using matrix-assisted laser desorption ionization-time of flight mass spectrometry. Curr Med Mycol 2017;3:21–25
    [Google Scholar]
  24. Zvezdanova ME, Escribano P, Ruiz A, Martínez-Jiménez MC, Peláez T et al. Increased species-assignment of filamentous fungi using MALDI-TOF MS coupled with a simplified sample processing and an in-house library. Med Mycol 2018; Feb 9
    [Google Scholar]
  25. Schurko AM, Mendoza L, Lévesque CA, Désaulniers NL, de Cock AW et al. A molecular phylogeny of Pythium insidiosum. Mycol Res 2003;107:537–544 [CrossRef]
    [Google Scholar]
  26. Presser JW, Goss EM. Environmental sampling reveals that Pythium insidiosum is ubiquitous and genetically diverse in North Central Florida. Med Myco 2015;53:674–683 [CrossRef]
    [Google Scholar]
  27. Vilela R, Montalva C, Luz C, Humber RA, Mendoza L. Pythium insidiosum isolated from infected mosquito larvae in central Brazil. Acta Trop 2018;185:344–348 [CrossRef]
    [Google Scholar]
  28. Nobrega de Almeida J, Del Negro GM, Grenfell RC, Vidal MS, Thomaz DY et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry for differentiation of the dimorphic fungal species Paracoccidioides brasiliensis and Paracoccidioides lutzii. J Clin Microbiol 2015;53:1383–1386 [CrossRef]
    [Google Scholar]
  29. Farmer AR, Murray CK, Driscoll IR, Wickes BL, Wiederhold N et al. Combat-Related Pythium aphanidermatum invasive wound infection: case report and discussion of utility of molecular diagnostics. J Clin Microbiol 2015;53:1968–1975 [CrossRef]
    [Google Scholar]
  30. Chindamporn A, Vilela R, Hoag KA, Mendoza L. Antibodies in the sera of host species with pythiosis recognize a variety of unique immunogens in geographically divergent Pythium insidiosum strains. Clin Vaccine Immunol 2009;16:330–336 [CrossRef]
    [Google Scholar]
  31. Kittichotirat W, Patumcharoenpol P, Rujirawat T, Lohnoo T, Yingyong W et al. Draft genome and sequence variant data of the oomycete Pythium insidiosum strain Pi45 from the phylogenetically-distinct Clade-III. Data Brief 2017;15:896–900 [CrossRef]
    [Google Scholar]
  32. Anderson NW, Buchan BW, Riebe KM, Parsons LN, Gnacinski S et al. Effects of solid-medium type on routine identification of bacterial isolates by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2012;50:1008–1013 [CrossRef]
    [Google Scholar]
  33. Wieme AD, Spitaels F, Aerts M, De Bruyne K, Van Landschoot A et al. Effects of growth medium on matrix-assisted laser Desorption–Ionization time of flight mass spectra: a case study of acetic acid bacteria. Appl Environ Microbiol 2014;80:1528–1538 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000941
Loading
/content/journal/jmm/10.1099/jmm.0.000941
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error