1887

Abstract

Free-living amoebae of the genus are the causative agents of severe sight-threatening infection of the cornea. This study was designed to characterize the genotype of 20 spp. isolates obtained from corneal scrapings of 183 suspected keratitis patients reporting to the Outpatient Department/Casualty Services of Dr R. P. Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India during the period 2011–2015. Corneal scrapings were inoculated onto 2 % non-nutrient agar plates overlaid with and incubated at 30 °C for 15 days. Amongst 183 suspected keratitis patients, 29 were found culture-positive for spp. out of which 20 samples were established in axenic culture for molecular analysis. DNA was isolated and PCR assay was performed for the amplification of the diagnostic fragment 3 (DF3) (∼280 bp) region of the 18S rRNA gene from axenic culture of 20 spp. isolates. genotyping was performed on the basis of variation in nucleotide sequences of the DF3 region of the 18S rRNA gene. In the phylogenetic analysis, 16 of the 20 isolates were found to be of prevalent genotype T4, two were of genotype T10 and the remaining two isolates were of unassigned genotypes. Hence, it was concluded that genotype T4 was found as the most predominant genotype involved in keratitis infections. Genotype T10, which had not been reported from India, was detected for the first time in two patients. Two isolates were found to be unique, which shared < 95 % homology with all the known genotypes (T1–T20) of spp.

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2016-05-01
2021-10-16
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References

  1. Arnalich-Montiel F., Lumbreras-Fernández B., Martín-Navarro C. M., 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 52:1213–1216 [View Article][PubMed]
    [Google Scholar]
  2. Bharathi J. M., Srinivasan M., Ramakrishnan R., Meenakshi R., Padmavathy S., Lalitha P. N. 2007; A study of the spectrum of Acanthamoeba keratitis: a three-year study at a tertiary eye care referral center in South India. Indian J Ophthalmol 55:37–42 [View Article][PubMed]
    [Google Scholar]
  3. Blaschitz M., Köhsler M., Aspöck H., Walochnik J. 2006; Detection of a serine proteinase gene in Acanthamoeba genotype T6 (Amoebozoa: Lobosea). Exp Parasitol 114:26–33 [View Article][PubMed]
    [Google Scholar]
  4. Booton G. C., Kelly D. J., Chu Y. W., Seal D. V., Houang E., Lam D. S., Byers T. J., Fuerst P. A. 2002; 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong. J Clin Microbiol 40:1621–1625 [View Article][PubMed]
    [Google Scholar]
  5. Booton G. C., Visvesvara G. S., Byers T. J., Kelly D. J., Fuerst P. A. 2005; Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol 43:1689–1693 [View Article][PubMed]
    [Google Scholar]
  6. Dart J. K., Saw V. P., Kilvington S. 2009; Acanthamoeba keratitis: diagnosis and treatment update 2009. Am J Ophthalmol 148:487–499, e2 [View Article][PubMed]
    [Google Scholar]
  7. Fuerst P. A., Booton G. C., Crary M. 2015; Phylogenetic analysis and the evolution of the 18S rRNA gene typing system of Acanthamoeba . J Eukaryot Microbiol 62:69–84 [View Article][PubMed]
    [Google Scholar]
  8. Gast R. J., Ledee D. R., Fuerst P. A., Byers T. J. 1996; Subgenus systematics of Acanthamoeba: four nuclear 18S rDNA sequence types. J Eukaryot Microbiol 43:498–504 [View Article][PubMed]
    [Google Scholar]
  9. Grün A. L., Stemplewitz B., Scheid P. 2014; First report of an Acanthamoeba genotype T13 isolate as etiological agent of a keratitis in humans. Parasitol Res 113:2395–2400 [View Article][PubMed]
    [Google Scholar]
  10. Khan N. A. 2009 Acanthamoeba – Biology and Pathogenesis Wymondham: Caister Academic Press;
    [Google Scholar]
  11. Lalitha P., Lin C. C., Srinivasan M., Mascarenhas J., Prajna N. V., Keenan J. D., McLeod S. D., Acharya N. R., Lietman T. M., Porco T. C. 2012; Acanthamoeba keratitis in South India: a longitudinal analysis of epidemics. Ophthalmic Epidemiol 19:111–115 [View Article][PubMed]
    [Google Scholar]
  12. Landell M. F., Salton J., Caumo K., Broetto L., Rott M. B. 2013; Isolation and genotyping of free-living environmental isolates of Acanthamoeba spp. from bromeliads in Southern Brazil. Exp Parasitol 134:290–294 [View Article][PubMed]
    [Google Scholar]
  13. Ledee D. R., Iovieno A., Miller D., Mandal N., Diaz M., Fell J., Fini M. E., Alfonso E. C. 2009; Molecular identification of T4 and T5 genotypes in isolates from Acanthamoeba keratitis patients. J Clin Microbiol 47:1458–1462 [View Article][PubMed]
    [Google Scholar]
  14. Lorenzo-Morales J., Khan N. A., Walochnik J. 2015; An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment. Parasite 22:10 [View Article][PubMed]
    [Google Scholar]
  15. Manikandan P., Bhaskar M., Revathy R., John R. K., Narendran V., Panneerselvam K. 2004; Acanthamoeba keratitis – a six year epidemiological review from a tertiary care eye hospital in South India. Indian J Med Microbiol 22:226–230[PubMed]
    [Google Scholar]
  16. Nazar M., Haghighi A., Niyyati M., Eftekhar M., Tahvildar-Biderouni F., Taghipour N., Abadi A., Nazemalhosseini Mojarad E., Athari A. 2011; Genotyping of Acanthamoeba isolated from water in recreational areas of Tehran, Iran. J Water Health 9:603–608 [View Article][PubMed]
    [Google Scholar]
  17. Nuprasert W., Putaporntip C., Pariyakanok L., Jongwutiwes S. 2010; Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol 48:4636–4640 [View Article][PubMed]
    [Google Scholar]
  18. Pasricha G., Sharma S., Garg P., Aggarwal R. K. 2003; Use of 18S rRNA gene-based PCR assay for diagnosis of Acanthamoeba keratitis in non-contact lens wearers in India. J Clin Microbiol 41:3206–3211 [View Article][PubMed]
    [Google Scholar]
  19. Patel A., Hammersmith K. 2008; Contact lens-related microbial keratitis: recent outbreaks. Curr Opin Ophthalmol 19:302–306 [View Article][PubMed]
    [Google Scholar]
  20. Prashanth K., Pasricha G., Sharma S. 2011; Fluorescence amplified fragment length polymorphism for subtyping of genotypes of Acanthamoeba isolated from patients with keratitis. Indian J Med Res 133:83–87[PubMed]
    [Google Scholar]
  21. Pussard M., Pons R. 1977; [Morphologies of the cystic wall and taxonomy of the genus Acanthamoeba (Protozoa, Amoebida)]. Protistologica (Paris) 13:557–610 (in French)
    [Google Scholar]
  22. Rahdar M., Niyyati M., Salehi M., Feghhi M., Makvandi M., Pourmehdi M., Farnia S. 2012; Isolation and genotyping of Acanthamoeba strains from environmental sources in Ahvaz City, Khuzestan province, southern Iran. Iran J Parasitol 7:22–26[PubMed]
    [Google Scholar]
  23. Schroeder J. M., Booton G. C., Hay J., Niszl I. A., Seal D. V., Markus M. B., Fuerst P. A., Byers T. J. 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 39:1903–1911 [View Article][PubMed]
    [Google Scholar]
  24. Sharifi N., Botero-Kleiven S., Ohman D., Barragan A., Winiecka-Krusnell J. 2010; Genotypic characterization of Acanthamoeba spp. causing ocular infections in Swedish patients: identification of the T15 genotype in a case of protracted keratitis. Scand J Infect Dis 42:781–786 [View Article][PubMed]
    [Google Scholar]
  25. Sharma S., Pasricha G., Das D., Aggarwal R. K. 2004; Acanthamoeba keratitis in non-contact lens wearers in India: DNA typing-based validation and a simple detection assay. Arch Ophthalmol 122:1430–1434 [View Article][PubMed]
    [Google Scholar]
  26. Sharma R., Jhanji V., Satpathy G., Sharma N., Khokhar S., Agarwal T. 2013; Coinfection with Acanthamoeba and Pseudomonas in contact lens-associated keratitis. Optom Vis Sci 90:e53–e55 [View Article][PubMed]
    [Google Scholar]
  27. Siddiqui R., Khan N. A. 2012; Biology and pathogenesis of Acanthamoeba . Parasit Vectors 5:6 [View Article][PubMed]
    [Google Scholar]
  28. Stothard D. R., Schroeder-Diedrich J. M., Awwad M. H., Gast R. J., Ledee D. R., Rodriguez-Zaragoza S., Dean C. L., Fuerst P. A., Byers T. J. 1998; The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45:45–54 [View Article][PubMed]
    [Google Scholar]
  29. Tamura K., Stecher G., Peterson D., Filipski A., Kumar S. 2013; mega6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30:2725–2729 [View Article][PubMed]
    [Google Scholar]
  30. Walochnik J., Scheikl U., Haller-Schober E. M. 2015; Twenty years of Acanthamoeba diagnostics in Austria. J Eukaryot Microbiol 62:3–11 [View Article][PubMed]
    [Google Scholar]
  31. Zhao G., Sun S., Zhao J., Xie L. 2010; Genotyping of Acanthamoeba isolates and clinical characteristics of patients with Acanthamoeba keratitis in China. J Med Microbiol 59:462–466 [View Article][PubMed]
    [Google Scholar]
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