title: Rapid detection of pathogenic leptospires by lyophilized reagent-based polymerase chain reaction.
abstract: A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed.   We report the development of a lyophilized reagent-based polymerase chain reaction (PCR)   assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine   the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the   sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able   to detect 10² and 10³ leptospires/ml in spiked urine and blood samples, respectively. In   experimentally infected animals, leptospiral DNA could be detected in blood and lung samples   as early as Day 1 post infection. This assay was also shown to be stable and remained   sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based   PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and   reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine.
authors.affiliation: Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 Serdang, Malaysia. svlee7@gmail.com
authors.forename: S V
authors.initials: SV
authors.lastname: Lee
authors.forename: E S
authors.initials: ES
authors.lastname: Tai
authors.forename: A R
authors.initials: AR
authors.lastname: Mutalib
authors.forename: S
authors.initials: S
authors.lastname: Khairani-Bejo
authors.forename: A R
authors.initials: AR
authors.lastname: Bahaman
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D001769:Blood / Q000382:microbiology; D006224:Cricetinae; D004195:Disease Models, Animal; D005612:Freeze Drying; D006801:Humans; D007919:Leptospira / Q000235:genetics* / Q000302:isolation & purification*; D007922:Leptospirosis / Q000175:diagnosis*; D008168:Lung / Q000382:microbiology; D008647:Mesocricetus; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014556:Urine / Q000382:microbiology
chemical_list: D011933:Reagent Kits, Diagnostic