title: Application of a loop-mediated isothermal amplification method for the detection of pathogenic Leptospira.
abstract: Leptospirosis is an emerging infectious disease, which is considered to be the most widespread zoonotic disease in the world. There are more than 230 known serovars in the genus Leptospira. A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of pathogenic Leptospira spp. was developed and evaluated through amplification of the lipL41 gene coding for the outer membrane protein LipL41. The LAMP assay did not rely on the isolation and culture of leptospires, and no cross-reactivity was observed with other bacterial species. A SYBR Green I-based LAMP assay was also carried out for the real-time detection of DNA amplification. The lower detection limit of the LAMP assay was approximately 100 copies, which was the same as the polymerase chain reaction (PCR) and real-time PCR assays. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis of the amplified product. The LAMP assay is easy to perform and inexpensive, and so may be applied in the rapid and specific diagnosis of Leptospira.
authors.affiliation: Department of Medical Microbiology and Parasitology, Zhejiang University, Hangzhou, China.
authors.forename: Xu'ai
authors.initials: X
authors.lastname: Lin
authors.forename: Yin
authors.initials: Y
authors.lastname: Chen
authors.forename: Yiyu
authors.initials: Y
authors.lastname: Lu
authors.forename: Juying
authors.initials: J
authors.lastname: Yan
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001483:Base Sequence; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D006801:Humans; D007919:Leptospira / Q000235:genetics / Q000302:isolation & purification*; D007922:Leptospirosis / Q000175:diagnosis*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
chemical_list: D001425:Bacterial Outer Membrane Proteins; D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I