title: Evaluation of loop-mediated isothermal amplification assay for rapid diagnosis ofAcanthamoebakeratitis.
abstract: BACKGROUND
  The clinical features of Acanthamoeba keratitis (AK) are non-specific and closely resemble bacterial, viral and fungal keratitis.
  
  
  METHODS
  We compared loop-mediated isothermal amplification (LAMP) with microscopy, non-nutrient agar (NNA) culture and polymerase chain reaction (PCR) in clinical suspects of AK.
  
  
  RESULTS
  Of 52 clinical samples (42 AK suspects and 10 proven bacterial, viral or fungal keratitis), 3 were positive by direct microscopy (sensitivity 60%, confidence interval [CI]: 17%-92.7%), and 5 by NNA culture, 18S rDNA PCR and LAMP (sensitivity 100%, CI: 46.3%-100%). The limit of detection of Acanthamoeba DNA was 1 pg/μl by both LAMP and PCR.
  
  
  CONCLUSIONS
  PCR and LAMP assays targeting 18S rDNA gene were found particularly suitable for a rapid and accurate diagnosis of AK. LAMP assay takes 2-3 h lesser than PCR, and thus offers a rapid, highly sensitive and specific, simple and affordable diagnostic modality for patients suspected of AK, especially in resource limited settings.
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Mewara
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Sumeeta
authors.initials: S
authors.lastname: Khurana
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Shakila
authors.initials: S
authors.lastname: Yoonus
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Kirti
authors.initials: K
authors.lastname: Megha
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Parveen
authors.initials: P
authors.lastname: Tanwar
authors.affiliation: Department of Ophthalmology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Gupta
authors.affiliation: Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Sehgal
mesh_terms: D000048:Acanthamoeba / Q000235:genetics / Q000302:isolation & purification*; D015823:Acanthamoeba Keratitis / Q000175:diagnosis*; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods; D008853:Microscopy / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012337:RNA, Ribosomal, 18S