Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지
DOI QR코드

DOI QR Code

Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba

  • Yang, Hye-Won (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Lee, Yu-Ran (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Inoue, Noboru (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine) ;
  • Jha, Bijay Kumar (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Sylvatrie Danne, Dinzouna-Boutamba (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Kim, Hong-Kyun (Department of Ophthalmology, Kyungpook National University Hospital) ;
  • Lee, Junhun (Department of Ophthalmology, Kyungpook National University Hospital) ;
  • Goo, Youn-Kyoung (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Kong, Hyun-Hee (Department of Parasitology, Dong-A University, College of Medicine) ;
  • Chung, Dong-Il (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Hong, Yeonchul (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine)
  • Received : 2013.04.19
  • Accepted : 2013.05.20
  • Published : 2013.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

Keywords

References

  1. Khan NA. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev 2006; 30: 564-595. https://doi.org/10.1111/j.1574-6976.2006.00023.x
  2. Corsaro D, Venditti D. Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa, Acanthamoebida). Parasitol Res 2010; 107: 233-238. https://doi.org/10.1007/s00436-010-1870-6
  3. Nuprasert W, Putaporntip C, Pariyakanok L, Jongwutiwes S. Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol 2010; 48: 4636-4640. https://doi.org/10.1128/JCM.01090-10
  4. Ledee DR, Hay J, Byers TJ, Seal DV, Kirkness CM. Acanthamoeba griffini. Molecular characterization of a new corneal pathogen. Invest Ophthalmol Vis Sci 1996; 37: 544-550.
  5. Walochnik J, Obwaller A, Aspock H. Correlations between morphological, molecular biological, and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. Appl Environ Microbiol 2000; 66: 4408-4413. https://doi.org/10.1128/AEM.66.10.4408-4413.2000
  6. Yu HS, Kong HH, Kim SY, Hahn YH, Hahn TW, Chung DI. Laboratory investigation of Acanthamoeba lugdunensis from patients with keratitis. Invest Ophthalmol Vis Sci 2004; 45: 1418-1426. https://doi.org/10.1167/iovs.03-0433
  7. Xuan YH, Chung BS, Hong YC, Kong HH, Hahn TW, Chung DI. Keratitis by Acanthamoeba triangularis: report of cases and characterization of isolates. Korean J Parasitol 2008; 46: 157-164. https://doi.org/10.3347/kjp.2008.46.3.157
  8. Dart JK, Saw VP, Kilvington S. Acanthamoeba keratitis: diagnosis and treatment update 2009. Am J Ophthalmol 2009; 148: 487-499 e482. https://doi.org/10.1016/j.ajo.2009.06.009
  9. Patel A, Hammersmith K. Contact lens-related microbial keratitis: recent outbreaks. Curr Opin Ophthalmol 2008; 19: 302-306. https://doi.org/10.1097/ICU.0b013e3283045e74
  10. Mozayan A, Madu A, Channa P. Laser in-situ keratomileusis infection: review and update of current practices. Curr Opin Ophthalmol 2011; 22: 233-237. https://doi.org/10.1097/ICU.0b013e3283479ed4
  11. Garg P, Chaurasia S, Vaddavalli PK, Muralidhar R, Mittal V, Gopinathan U. Microbial keratitis after LASIK. J Refract Surg 2010; 26: 209-216. https://doi.org/10.3928/1081597X-20100224-07
  12. Panjwani N. Pathogenesis of Acanthamoeba keratitis. Ocul Surf 2010; 8: 70-79. https://doi.org/10.1016/S1542-0124(12)70071-X
  13. Hammersmith KM. Diagnosis and management of Acanthamoeba keratitis. Curr Opin Ophthalmol 2006; 17: 327-331. https://doi.org/10.1097/01.icu.0000233949.56229.7d
  14. Mathers WD, Nelson SE, Lane JL, Wilson ME, Allen RC, Folberg R. Confirmation of confocal microscopy diagnosis of Acanthamoeba keratitis using polymerase chain reaction analysis. Arch Ophthalmol 2000; 118: 178-183. https://doi.org/10.1001/archopht.118.2.178
  15. Schroeder JM, Booton GC, Hay J, Niszl IA, Seal DV, Markus MB, Fuerst PA, Byers TJ. 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 2001; 39: 1903-1911. https://doi.org/10.1128/JCM.39.5.1903-1911.2001
  16. Sharma S, Srinivasan M, George C. Acanthamoeba keratitis in non-contact lens wearers. Arch Ophthalmol 1990; 108: 676-678. https://doi.org/10.1001/archopht.1990.01070070062035
  17. Srinivasan M, Burman S, George C, Nirmalan PK. Non-contact lens related Acanthamoeba keratitis at a tertiary eye care center in south India: Implications for eye care programs in the region. Med Sci Monit 2003; 9: CR125-129.
  18. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 2000; 28: E63. https://doi.org/10.1093/nar/28.12.e63
  19. Mori Y, Notomi T. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemother 2009; 15: 62-69. https://doi.org/10.1007/s10156-009-0669-9
  20. Mori Y, Nagamine K, Tomita N, Notomi T. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 2001; 289: 150-154. https://doi.org/10.1006/bbrc.2001.5921
  21. Goto M, Honda E, Ogura A, Nomoto A, Hanaki K. Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques 2009; 46: 167-172. https://doi.org/10.2144/000113072
  22. Tomita N, Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 2008; 3: 877-882. https://doi.org/10.1038/nprot.2008.57
  23. Lek-Uthai U, Passara R, Roongruangchai K, Buddhirakkul P, Thammapalerd N. Rapid identification of Acanthamoeba from contact lens case using loop-mediated isothermal amplification method. Exp Parasitol 2009; 121: 342-345. https://doi.org/10.1016/j.exppara.2008.12.009
  24. Kong HH, Shin JY, Yu HS, Kim J, Hahn TW, Hahn YH, Chung DI. Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J Clin Microbiol 2002; 40: 1199-1206. https://doi.org/10.1128/JCM.40.4.1199-1206.2002
  25. Boggild AK, Martin DS, Lee TY, Yu B, Low DE. Laboratory diagnosis of amoebic keratitis: comparison of 4 diagnostic methods for different types of clinical specimens. J Clin Microbiol 2009; 47: 1314-1318. https://doi.org/10.1128/JCM.00173-09
  26. Enomoto Y, Yoshikawa T, Ihira M, Akimoto S, Miyake F, Usui C, Suga S, Suzuki K, Kawana T, Nishiyama Y, Asano Y. Rapid diagnosis of herpes simplex virus infection by a loop-mediated isothermal amplification method. J Clin Microbiol 2005; 43: 951-955. https://doi.org/10.1128/JCM.43.2.951-955.2005
  27. Kaneko H, Kawana T, Fukushima E, Suzutani T. Tolerance of loop-mediated isothermal amplification to a culture medium and biological substances. J Biochem Biophys Methods. 2007; 70: 499-501. https://doi.org/10.1016/j.jbbm.2006.08.008
  28. Booton GC, Visvesvara GS, Byers TJ, Kelly DJ, Fuerst PA. Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol 2005; 43: 1689-1693. https://doi.org/10.1128/JCM.43.4.1689-1693.2005

Cited by

  1. Molecular diagnosis in clinical parasitology: When and why? vol.239, pp.11, 2013, https://doi.org/10.1177/1535370214523880
  2. Acanthamoeba keratitis: improving the Scottish diagnostic service for the rapid molecular detection of Acanthamoeba species vol.64, pp.7, 2015, https://doi.org/10.1099/jmm.0.000086
  3. Development of Visually Improved Loop Mediated Isothermal Amplification for the Diagnosis of Plasmodium vivax Malaria in a Tertiary Hospital in Chandigarh, North India vol.98, pp.5, 2018, https://doi.org/10.4269/ajtmh.17-0857
  4. Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR vol.54, pp.2, 2018, https://doi.org/10.7845/kjm.2018.8012
  5. Surveillance of viable Acanthamoeba spp. and Naegleria fowleri in major water sources for tap water in Korea vol.54, pp.3, 2018, https://doi.org/10.7845/kjm.2018.8040
  6. A simple and visible detection method for the rapid diagnosis of Ustilaginoidea virens in rice seeds by a loop‐mediated isothermal amplification assay vol.169, pp.6, 2021, https://doi.org/10.1111/jph.12991
  7. Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample vol.11, pp.1, 2013, https://doi.org/10.1038/s41598-021-00968-2