title: A field-expedient method for detection of leptospirosis causative agents in rodents.
abstract: We have developed a thermal-stable, pathogenic Leptospira TaqMan PCR assay intended to support pathogen surveillance in reservoir populations. The assay is packaged specifically for use with a portable, ruggedized, real-time PCR thermocycler. Limit of detection was established at ≤ 100 fg (20 organisms). Sensitivity and specificity were 100% concordant with conventional PCR results using a broad test panel of human pathogenic and nonpathogenic Leptospira, genetic near neighbors, and clinically significant organisms. In blind testing using a panel (n=50) of pathogenic Leptospira infected and noninfected Rattus species samples, assay sensitivity results were 100% concordant with conventional PCR. Tests performed under field conditions using wild-collected rodent kidney extracts demonstrated the mobility of the system. During field evaluation, samples were processed and analyzed in 3 hours. Thermal stabilized reagents allowed transportation, storage, and analyses under ambient temperatures. The system provides a promising aid in leptospirosis control programs.
authors: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
authors: James C
authors: JC
authors: McAvin
authors: Ampornpan
authors: A
authors: Kengluecha
authors: Ratree
authors: R
authors: Takhampunya
authors: Jason H
authors: JH
authors: Richardson
mesh_terms: D000818:Animals; D004197:Disease Reservoirs / Q000382:microbiology / Q000662:veterinary; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000472:pathogenicity; D007922:Leptospirosis / Q000175:diagnosis* / Q000517:prevention & control; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012376:Rodent Diseases / Q000382:microbiology; D012377:Rodentia; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods