Rapid point ‐of‐care diagnostics for the detection of Chlamydia pecorum in koalas (Phascolarctos cinereus) using loop‐mediated isothermal amplification without nucleic acid purification

In this study, we describe the development and evaluation of aChlamydia pecorum‐specific loop‐mediated isothermal amplification (LAMP) assay on the Genie ® III as a rapid point‐of care diagnostic test that can be easily incorporated into wildlife hospitals or koala field surveys. Furthermore, we present a simplistic one‐step lysis method for template preparation using koala conjunctival, urogenital, and rectal swabs. We have demonstrated theC. pecorum LAMP assay is highly sensitive and specific compared with real ‐time PCR, with a level of detection of 44C. pecorum IFU/ml. AbstractInfectious disease, predominately chlamydiosis, contributes significantly to the decline in health of wild koala (Phascolarctos cinereus) populations in some regions of Australia. In this study, we describe the development and evaluation of a simple, sensitive, and specific loop ‐mediated isothermal amplification (LAMP) assay for the detection ofChlamydia pecorum in koalas as a point ‐of‐care diagnostic tool that can be used in any wildlife hospital and in the field on specialized instrumentation. A set of primers targeting a 188‐bp region of theC.  pecorum genome was designed. 100% specificity of the LAMP assay was revealed by demonstrating no cross ‐reactivity with 33 nontarget pathogens, and complete correlation with qPCR results for 43 clinical swabs collected opportunistically from wildlife hospitals. In sensitivity evaluations, the technique successfully detected serial d...
Source: MicrobiologyOpen - Category: Microbiology Authors: Tags: ORIGINAL ARTICLE Source Type: research