title: Development of loop-mediated isothermal amplification method for detecting Wuchereria bancrofti DNA in human blood and vector mosquitoes.
abstract: We have developed loop-mediated isothermal amplification (LAMP) method to detect Wuchereria bancrofti DNA. The sensitivity and specificity of LAMP method were equivalent to those of PCR method which detects SspI repeat sequence in W. bancrofti genomic DNA: both methods detected one thousandth of W. bancrofti DNA from one microfilaria (Mf), and did not cross-react with DNAs of Brugia malayi, B. pahangi, Dirofilaria immitis, human and Culex quinquefasciatus. We also examined the sensitivity of LAMP using the mimic samples of patient's blood or blood-fed mosquitoes containing one W. bancrofti Mf per sample. The LAMP method was able to detect W. bancrofti DNA in 1000 μl of blood or in a pool of 60 mosquitoes, indicating its usefulness in detecting/monitoring W. bancrofti infection in humans and vector mosquitoes in endemic areas.
authors.affiliation: Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan. htakagi@aichi-med-u.ac.jp
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Takagi
authors.forename: Makoto
authors.initials: M
authors.lastname: Itoh
authors.forename: Shinji
authors.initials: S
authors.lastname: Kasai
authors.forename: Thishan C
authors.initials: TC
authors.lastname: Yahathugoda
authors.forename: Mirani V
authors.initials: MV
authors.lastname: Weerasooriya
authors.forename: Eisaku
authors.initials: E
authors.lastname: Kimura
mesh_terms: D000818:Animals; D001483:Base Sequence; D002009:Brugia / Q000235:genetics; D003465:Culex / Q000235:genetics; D018509:DNA, Helminth / Q000032:analysis / Q000235:genetics*; D004182:Dirofilaria / Q000235:genetics; D005368:Filariasis / Q000097:blood / Q000175:diagnosis / Q000469:parasitology; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D012680:Sensitivity and Specificity; D014958:Wuchereria bancrofti / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D018509:DNA, Helminth