title: Rapid identification of Acanthamoeba from contact lens case using loop-mediated isothermal amplification method.
abstract: A method employing loop-mediated isothermal amplification (LAMP) of 18S ribosomal RNA gene was developed to detect Acanthamoeba in contact lens cases. A prevalence of 7% (10/150) was detected, with 100% sensitivity and 100% specificity when compared with the standard culture technique. Using visual inspection of turbidity a minimum of 10pg of Acanthamoeba DNA could be detected, 10 times more sensitive than quantitative PCR employing two of the LAMP primers. The production of LAMP amplicons was confirmed by gel-electrophoresis and ethidium bromide staining. The LAMP procedure takes less than 2h to perform and will be useful for incorporation into a point-of-care screening of suspected Acanthamoeba infection.
authors.affiliation: Department of Parasitology, Faculty of Public Health, Mahidol University, 420/1 Rajavithi Road, Rajathewee District, Bangkok 10400, Thailand. phulu@mahidol.ac.th
authors.forename: Usa
authors.initials: U
authors.lastname: Lek-Uthai
authors.forename: Rangsima
authors.initials: R
authors.lastname: Passara
authors.forename: Kosol
authors.initials: K
authors.lastname: Roongruangchai
authors.forename: Prayute
authors.initials: P
authors.lastname: Buddhirakkul
authors.forename: Nitaya
authors.initials: N
authors.lastname: Thammapalerd
mesh_terms: D000048:Acanthamoeba / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D003261:Contact Lenses / Q000469:parasitology*; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S