Abstract
We developed an assay comprising crude DNA lysis by simple heat treatment coupled loop-mediated isothermal amplification with hydroxynaphthol blue for Chlamydia trachomatis detection (petty patent pending), and evaluated the developed assay for its feasibility as a one-step point-of-care detection on 284 endocervical swab specimens from clinically symptomatic C.trachomatis and healthy subjects. This assay is sensitive to 004pg of ompA, specific with six primers targeting C.trachomatis ompA region, rapid (45min total assay time), inexpensive (approx. 3USD/reaction), does not require sophisticated instrumentation, and has comparable assay effectiveness (95% specificity, 90-100% sensitivity) to bacterial DNA isolation by a commercial kit coupled with polymerase chain reaction and gel electrophoresis (98-100% specificity, 87-100% sensitivity) based on the clinical samples test. The test result could be read by naked eye through the colour change from violet (negative) to sky blue (positive) for C.trachomatis-infected specimens. Further, this assay uses all safe chemical reagents and is hence safe to the users.
