Development of DuplexPolymerase Chain Reactionin-house assay to Detect of Chlamydia trachomatis and Neisseria gonorrhoeaefrom Genital Discharge in Patient with Sexual Transmitted Infections

Wardoyo, E Hagni (2017) Development of DuplexPolymerase Chain Reactionin-house assay to Detect of Chlamydia trachomatis and Neisseria gonorrhoeaefrom Genital Discharge in Patient with Sexual Transmitted Infections. The Indonesian Journal of Biotechnology Medicine, 6 (1). pp. 53-65. ISSN 2301-5810

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Abstract

The incidence of sexually transmitted infections (STIs) with discharge in Indonesia to high-risk groups provide dominant pattern of Chlamydia trachomatis(CT) and Neisseriagonorrheae(NG) infection. Pathogenesis change of CT and NGinfection suggeststhat routine syndromic approach diagnostic is no longer accurate. To develop detection system of CTand NG using duplex polymerase chain reaction (PCR) assay to genital discharge in patient with STIs. Three phases: firstly was PCR assay optimalization to annealing time and temperature, primer concentration, centrifugation time and elution volume, secondly, specificity test and thirdly duplex PCR assay application to clinical specimen. Duplex PCR assay optimalization gave results as follow: annealing temperature and time 54°C and 60 seconds, CTand NG primer concentration were 0,7μM and0,5μM, centrifugation time 10 minutes, and elution volume 60 μl. Detection limit of duplex PCR to CTand NG 0.927 and 1.19 pg / PCR reaction, respectively. Duplex PCR to detect NG has ensitivity, specificity, positive predictive value and negative predictive value were 100%, 61.9%, 20%, and 100% in endocervical specimens, respectively and 75%, 40%, 50%, and 66.67%, in male urethral specimens respectively. Duplex PCR to detect CT was compared with chlamydial antigen detection test were show positive results higher both in endocervical and male urethral specimens (10:3 and 1:0). Development of detection system of CT and NG using duplex PCR assay is applicable to endocervical and male urethral swab specimens.

Item Type: Article
Keywords (Kata Kunci): sexual transmitted infections (STIs), genital discharge, Duplex PCR in-house assay,Chlamydia trachomatis, Neisseria gonorrhoeae
Subjects: R Medicine > R Medicine (General)
Divisions: Fakultas Kedokteran
Depositing User: dr.Sp.MK Eustachius Hagni Wardoyo
Date Deposited: 14 Nov 2018 02:23
Last Modified: 14 Nov 2018 02:23
URI: http://eprints.unram.ac.id/id/eprint/9981

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