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Objective: Currently, Chlamydia trachomatis (Chlamydia trachomatis, CT) detection method based on fluorescence quantitative Taqman probe technique PCR, the method has high sensitivity and specificity and good and widely used in clinical. However, Taqman probe technology has the following two problems: First, because the probe labeled at both ends of the fluorescent dye, so the higher cost of synthesis and purification; Second, the detection process requires three steps: denaturation, hybridization, extension hydrolysis, detection time than long. Dimer scorpion probes were labeled fluorescent dyes in two complementary single-stranded DNA, from principle to overcome the Taqman probe labeling and purification difficult to reduce the cost and other shortcomings; eliminating the step of detecting the enzymatic hydrolysis process, can shorten the detection time. This study intends to establish a rapid, accurate, inexpensive, easy to operate, as a dimer scorpion probe quantitative PCR-based method for detecting CT. Methods: gene cloning technology, the CT conserved sequence major outer membrane protein gene (MOMP) was cloned into pMD18-T vector to construct a recombinant plasmid DNA standards. According MOMP gene sequence dimer scorpion probe, optimizing quantitative PCR system. Using four standard series and a positive specimens, its coefficient of variation (CV) and inter-assay coefficients of variation were determined; using Chlamydia pneumoniae, Chlamydia psittaci, 15 standard serotype CT, and 16 kinds of common genital tract pathogens its specificity was determined; using this detection system with two kinds of FDA-approved molecular diagnostic methods, respectively, of 148 cases of clinical specimens for testing, according to the CT diagnosis of the \sentenced for the specimen to specimen CT infection, the detection method to evaluate the performance of clinical testing. Results: 1) the recombinant plasmid. 2) to establish a type of fluorescent probes based on dimer scorpion fluorescence quantitative PCR method, the linear range of 25 ~ 1010copies / reaction tube; different concentrations of samples intra-assay CV range of 4.92% ~ 21.73%, inter-assay CV range 6.94% ~ 27.17%; specificity was 100%. 3) preliminary clinical applications show that the method can accurately diagnose 98.6% once the clinical diagnosis of specimens, is a fast, easy to operate for the molecular diagnosis of Chlamydia trachomatis infection in new ways. Conclusion: the establishment of a fluorescent probe technology dimer scorpion as a platform for Chlamydia trachomatis fluorescence quantitative polymerase chain reaction (FQ-PCR) method. The established method for optimizing and methodological evaluation, research shows that the method is reliable and fast detection, specificity high sensitivity, low reagent costs than benefits.
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