DEVELOPMENT AND VALIDATION OF AN IN-HOUSE REAL-TIME PCR ASSAY FOR THE DETECTION OF CHLAMYDIA TRACHOMATIS IN SEXUALLY TRANSMITTED INFECTIONS CLINIC
DOI:
https://doi.org/10.14456/taj.2020.3Keywords:
Chlamydia trachomatis, Polymerase Chain Reaction (PCR), In-house real-time PCRAbstract
In providing laboratory diagnostic service for Chlamydia trachomatis (C. trachomatis), there is a need to develop an accurate and affordable test kit. This study aimed to assess efficiency and cost of in-house real-time PCR assay as compare to commercial real-time PCR test kit in the diagnosis of C. trachomatis infection in routine service sexually transmitted infections (STIs) laboratory. In this study, known 100 positive and 30 negative samples for C. trachomatis as identified by cell culture, a standard method in diagnosis of C. trachomatis, were used in the comparison. All 130 samples were tested by in-house real-time
PCR in which primer and probe was developed to specify with C. trachomatis at cryptic plasmid, and commercial test kit (Anyplex CT/NG real-time detection (V3.1) kit, Gene Excellence Co. LTD., South Korea). Sensitivity, specificity, positive and negative predictive value were analyzed both for in-house real-time PCR and commercial kit against cell culture. Costs of the two tests were compared.
Results showed that tests performed by in-house real time PCR was accurate and comparable with commercial real time PCR kit. The sensitivity, specificity, and positive and negative predictive values of the in-house real time PCR were 100%, 96.7%, 99% and 100%, respectively. Cost of the in-house real-time PCR was 177 baht/test while commercial kit was 808 baht/test.
One of the samples negative for C. trachomatis by cell-culture was found to be positive both by in-house real-time PCR and commercial kit. It is possible that real-time PCR technique may have higher sensitivity value over cell culture method. Based on the finding from this study, it can be concluded that in house real time PCR has excellent accuracy similar to commercial real-time PCR kit but with lower cost. The in-house real-time PCR for C. trachomatis detection kit can be used in routine service STIs Laboratory.
Downloads
References
2. Orellana MA, Gómez-Lus ML, Lora D. Sensitivity of Gram stain in the diagnosis of urethritis in men. Sexually Transmitted Infections.2012, 88(4):284-87.
3. Lai-King Ng, Irene E Martin. The laboratory diagnosis of Neisseria gonorrhoeae. Can J Infect Dis Med Microbiol 2005; 16(1): 15-25.
4. Kanupriya G, LaDraka B, Rakesh KB, Christen GP, Xiaofei C, Rachel JG, et al. Performance of Chlamydia trachomatis OmcB Enzyme-Linked Immunosorbent Assay in Serodiagnosis of Chlamydia trachomatis Infection in Women. Journal of clinical microbiology. 2018, 56(9): 275-18.
5. Bachmann LH et al. Nucleic acid amplification tests for diagnosis of Neisseria gonorrhoeae and Chlamydia trachomatis rectal infections. Journal of Clinical Microbiology 2010, 48(5): 1827-32.
6. Schachter J et al. Nucleic acid amplification tests in the diagnosis of Chlamydial and gonococcal infections of the oropharynx and rectum in men who have sex with men. Sexually transmitted diseases 2008, 35(7): 637-42.
7. Manisha G, Anima S, Shravan KM. Molecular Diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae in Women at High Risk of Sexually Transmitted Infections. J Microb Biochem Technol 2017, 9(6): 321-24.
8. Hopkins MJ, Ashton J, Alloba F, et al. Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine. Sex Transm Infect 2010, 86: 207-11.
