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Design of Chlamydia Trachomatis Diagnostic Kit in Symptomatic Women Using Real Time PCR
Hamideh Rouhani nejad 1, Negar Rahmati2 , Fahimeh Nemati mansor3 , Darya Amiri4
1- Malek-Ashtar University of Technology, Tehran, Iran , rohaninejhad@gmail.com
2- Department of Microbioology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
3- 3. Department of biotechnology, Faculty of Science and Technologies, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran
4- Malek-Ashtar University of Technology, Tehran, Iran
Abstract:   (71 Views)
Background & Objective: Chlamydia trachomatis is one of the most common sexually transmitted diseases worldwide. Screening women for Chlamydia trachomatis in developing countries is highly desirable due to the ineffectiveness of the available diagnostic methods. The aim of this study is to design a detection kit for Chlamydia trachomatis in symptomatic women.
Materials & Methods: A total of 50 clinical specimens of symptomatic women were collected. The OMP1 gene sequence was extracted from the gene bank and the primer and probe were designed appropriately by Mega6 software. The extracted DNA was amplified by PCR. In order to provide positive control, PCR product was cloned into Pjet1.2 vector. To determine the sensitivity, DNA dilution was performed. To determine the specificity, DNA of vaginal bacteria was extracted and Real Time PCR was done.
Results: In this study, we designed a kits with 100% specificity and 10pg/ϥl sensitivity in detecting chlamydial infection in symptomatic women.
Conclusion: With the use of designed primers and real time, we can detect 100% of chlamydia infections.
 
Keywords: Real Time PCR, Chlamydia trachomatis, OMP1 gene
     
Type of Study: Research | Subject: Microbiology
Received: 2019/07/6 | Accepted: 2019/08/31
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