Skip to main content

Advertisement

SpringerLink
Log in

A simple PCR condition for detection of a single cyst of Acanthamoeba species

  • Short Communication
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Acanthamoeba is a free-living protozoan with a worldwide distribution in a variety of natural and artificial habitats. It has even been found in contact lens solution. Acanthamoeba spp. can cause infections such as granulomatous amoebic encephalitis and amoebic keratitis. Specific and sensitive diagnosis of Acanthamoeba infections can prevent clinical symptoms from worsening. Recently, PCR technique has been used for Acanthamoeba diagnosis. Unfortunately the dormant cyst of Acanthamoeba is resistant to chemical reagents; thus, most extraction of DNA uses a commercial DNA extraction kit for obtaining DNA for further use in polymerase chain reaction (PCR). Therefore in the present study, we improved the ability to diagnose Acanthamoeba using a simplified PCR technique. Interestingly, heating at 94°C for 10 min could release DNA which is amplified with specific primers designed from 16S rRNA. The PCR product is about 180 bp. This technique is a simple and efficient method for detection of Acanthamoeba—even a single cyst—and does not require high-cost reagents or complicated procedures to extract DNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

References

  • Bacon AS, Frazer DG, Dart JKG, Matheson M, Ficker LA, Wright P (1993) A review of 72 consecutive cases of Acanthamoeba keratitis, 1984–1992. Eye 7:719–725

    Article  PubMed  Google Scholar 

  • Beattie TK, Seal DV, Tomlinson A, McFadyen AK, Grimason AM (2003) Determination of amoebicidal activities of multipurpose contact lens solutions by using a most probable number enumeration technique. J Clin Microbiol 41:2992–3000

    Article  PubMed  CAS  Google Scholar 

  • Booton GC, Kelly DJ, Chu YW, Seal DV, Houang E, Lam DSC, Byers TJ, Fuerst PA (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

    Article  PubMed  CAS  Google Scholar 

  • Chang PC, Soong HK (1991) Acanthamoeba keratitis in non-contact lens wearers. Arch Ophthalmol 109:463–464

    PubMed  CAS  Google Scholar 

  • De Jonckheere J, Van de Voorde H (1976) Differences in destruction of cysts of pathogenic and nonpathogenic Naegleria and Acanthamoeba by chlorine. Appl Environ Microbiol 31:294–297

    PubMed  Google Scholar 

  • Di Cave D, Monno R, Bottalico P, Guerriero S, D’Amelio S, D’Orazi C, Berrilli F (2009) Acanthamoeba T4 and T15 genotypes associated with keratitis infections in Italy. Eur J Clin Microbiol Infect Dis 28:607–612

    Article  PubMed  Google Scholar 

  • Duguid IGM, Dart JKG, Morlet N, Allan BDS, Matheson M, Ficker L, Tuft S (1997) Outcome of Acanthamoeba keratitis treated with polyhexamethyl biguanide and propamidine. Ophthalmology 104:1587–1592

    PubMed  CAS  Google Scholar 

  • Foreman O, Sykes J, Ball L, Yang N, De Cock H (2004) Disseminated infection with Balamuthia mandrillaris in a dog. Vet Pathol 41:506–510

    Article  PubMed  CAS  Google Scholar 

  • Goldschmidt P, Rostane H, Saint-Jean C, Batellier L, Alouch C, Zito E, Bourcier T, Laroche L, Chaumeil C (2006) Effects of topical anaesthetics and fluorescein on the real-time PCR used for the diagnosis of Herpesviruses and Acanthamoeba keratitis. Br J Ophthalmol 90:1354–1356

    Article  PubMed  CAS  Google Scholar 

  • Hiti K, Walochnik J, Haller-Schober EM, Faschinger C, Aspöck H (2006) Efficacy of contact lens storage solutions against different Acanthamoeba strains. Cornea 25:423–427

    Article  PubMed  Google Scholar 

  • Iovieno A, Miller D, Lonnen J, Kilvington S, Alfonso EC (2011) Extraction of Acanthamoeba DNA by use of Chelex resin. J Clin Microbiol 49:476–477

    Article  PubMed  Google Scholar 

  • Johnston SP, Sriram R, Qvarnstrom Y, Roy S, Verani J, Yoder J, Lorick S, Roberts J, Beach MJ, Visvesvara G (2009) Resistance of Acanthamoeba cysts to disinfection in multiple contact lens solutions. J Clin Microbiol 47:2040–2045

    Article  PubMed  Google Scholar 

  • Jones DB, Visvesvara GS, Robinson NM (1975) Acanthamoeba polyphaga keratitis and Acanthamoeba uveitis associated with fatal meningoencephalitis. Trans Opthalmol Soc U K 95:221–232

    CAS  Google Scholar 

  • Khunkitti W, Lloyd D, Furr JR, Russell AD (1998) Acanthamoeba castellanii: growth, encystment, excystment and biocide susceptibility. J Infect 36:43–48

    Article  PubMed  CAS  Google Scholar 

  • Kilvington S, Anger C (2001) A comparison of cyst age and assay method of the efficacy of contact lens disinfectants against Acanthamoeba. Br J Ophthalmol 85:336–340

    Article  PubMed  CAS  Google Scholar 

  • Kosrirukvongs P, Wanachiwanawin D, Visvesvara GS (1999) Treatment of Acanthamoeba keratitis with chlorhexidine. Ophthalmology 106:798–802

    Article  PubMed  CAS  Google Scholar 

  • Lehmann OJ, Green SM, Morlet N, Kilvington S, Keys MF, Matheson MM, Dart JK, McGill JI, Watt PJ (1998) Polymerase chain reaction analysis of corneal epithelial and tear samples in the diagnosis of Acanthamoeba keratitis. Invest Ophthalmol Vis Sci 39:1961–1965

    Google Scholar 

  • Lorenzo-Morales J, López-Darias M, Martínez-Carretero E, Valladares B (2007) Isolation of potentially pathogenic strains of Acanthamoeba in wild squirrels from the Canary Islands and Morocco. Exp Parasitol 117:74–79

    Article  PubMed  CAS  Google Scholar 

  • Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16:273–307

    Article  PubMed  Google Scholar 

  • Marciano-Cabral F, Puffenbarger R, Cabral GA (2000) The increasing importance of Acanthamoeba infections. J Eukaryot Microbiol 47:29–36

    Article  PubMed  CAS  Google Scholar 

  • Marciano-Cabral F, MacLean R, Mensah A, Lapat-Polasko L (2003) Identification of Naegleria fowleri in domestic water sources by nested PCR. Appl Environ Microbiol 10(69):5864–5869

    Article  Google Scholar 

  • Mathers WD, Sutphin JE, Folberg R, Meier PA, Wenzel RP, Elgin RG (1996) Outbreak of keratitis presumed to be caused by Acanthamoeba. Am J Ophthalmol 121:129–142

    PubMed  CAS  Google Scholar 

  • Niszl IA, Markus MB (1998) Anti-Acanthamoeba activity of contact lens solutions. Br J Ophthalmol 82:1033–1038

    Article  PubMed  CAS  Google Scholar 

  • Pasricha G, Sharma S, Garg P, Aggarwal RK (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

    Article  PubMed  CAS  Google Scholar 

  • Preechawat P, Ratananikom U, Lerdvitayasakul R, Kunavisarut S (2007) Contact lens-related microbial keratitis. J Med Assoc Thai 90:737–743

    PubMed  Google Scholar 

  • Radford CF, Minassian DC, Dart JKG (2002) Acanthamoeba keratitis in England and Wales: incidence, outcome, and risk factors. Br J Ophthalmol 86:536–542

    Article  PubMed  CAS  Google Scholar 

  • Seal DV, Bron AJ, Hay J (1998) Ocular infection: investigation and treatment in practice. Community Eye Health 11(27):42

    Google Scholar 

  • Seitzman GD, Cevallos V, Margolis TP (2006) Rose bengal and lissamine green inhibit detection of herpes simplex virus by PCR. Am J Ophthalmol 141:756–758

    Article  PubMed  CAS  Google Scholar 

  • Sharma S, Srinivasan M, George C (1990) Acanthamoeba keratitis in non-contact lens wearers. Arch Ophthalmol 108:676–678

    Article  PubMed  CAS  Google Scholar 

  • Stehr-Green JK, Bailey TM, Brandt FH, Carr JH, Bond WW, Visvesvara GS (1987) Acanthamoeba keratitis in soft contact lens wearers. JAMA 258:57–60

    Article  PubMed  CAS  Google Scholar 

  • Yagi S, Schuster FL, Visvesvara GS (2008) Demonstration of Balamuthia and Acanthamoeba mitochondrial DNA in sectioned archival brain and other tissues by the polymerase chain reaction. Parasitol Res 102:491–497

    Article  PubMed  Google Scholar 

  • Zhang Y, Sun X, Wang Z, Li R, Luo S, Jin X, Deng S, Chen W (2004) Identification of 18S ribosomal DNA genotype of Acanthamoeba from patients with keratitis in north China. Invest Ophthalmol Vis Sci 45:1904–1907

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported in part by grants from: the Research Foundation of the Faculty of Medicine, Khon Kaen University; the Commission on Higher Education; and the Thailand Research Fund (TRG5480003).

Author information

Authors and Affiliations

  1. Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Porntip Laummaunwai, Wipaporn Ruangjirachuporn & Thidarut Boonmars

Authors
  1. Porntip Laummaunwai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wipaporn Ruangjirachuporn
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Thidarut Boonmars
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Thidarut Boonmars.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Laummaunwai, P., Ruangjirachuporn, W. & Boonmars, T. A simple PCR condition for detection of a single cyst of Acanthamoeba species. Parasitol Res 110, 1569–1572 (2012). https://doi.org/10.1007/s00436-011-2662-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-011-2662-3

Keywords

Search

Navigation