P30-01ACCURATE AND SPECIFIC DETECTION OF 13 GENOTYPES ASSOCIATED WITH SEXUALLY TRANSMITTED DISEASE BY PNA MEDIATED REAL TIME PCR

Sexually transmitted diseases (STD), also referred to as venereal diseases (VD) are diseases that can be spread by sexual activity or sexual contactless infection such as transmission via infected blood. Some of STD pathogens such as Gardnerella vaginalis (GV) show no symptoms in men but has the ability to infect women. Other pathogens such as Chlamydia trachomatis (CT) and Mycoplasma genitalium (MG) may appear chronic pain, itching, small fluid-filled blisters and painful sexual intercourse or even infertility in women and it can be prescribed antibiotics. Neisseria gonorrhea (NG) causes symptoms of cervicitis, urethritis and others in women and men both. NG or CT can be transmitted from a pregnant mother to unborn child. Therefore, detection of STD pathogens is essential for accurate treatment because each type of STD pathogens requires different prescription of antibiotics.

We have developed a highly sensitive and simple real-time PCR method to detect 13 types of STD pathogen from DNA in patient’s specimens such as cervical swab and urine both for female and male. This method (PANA RealTyper^TM) uses specific peptide nucleic acid (PNA) probe conjugated with a fluorescent dye and a quencher. These PNA probes are designed to their specific target sequences, which results in each having unique Tm value. 

Test was performed to obtain 40 cervical swab samples and 35 urine samples. DNA was extracted from 40 cervical swab samples and 35 urine samples. PANA RealTyper^TM STD analysis showed a concordance rate of 92% in 69 samples of 75 samples. Mismatched 6 samples were confirmed via sequencing, it showed that the result of PANA RealTyper^TM STD matched sequencing PCR. 40 of 85 clinical specimens were infected samples with two or more STD pathogens. 

RealTyper^TM STD was able to detect multiple pathogen targets at the same time. PANA RealTyper^TM STD is able to detect 13 types of different pathogens with detection limits as low as 5x10¹copies/rxn using standard materials. It provides genotyping of a total 13 types (Chlamydia trachomatis, Trichomonas vaginalis, Ureaplasma pavum, Ureaplasma urealyticum, Mycoplasma genitalium, Mycoplasma hominis, Neisseria gonorrhoeae, Haemophilus ducreyi, Herpes Simplex Virus 1, 2, Gardnerella Vaginalis, Candida albicans, Treponema pallidum) by real-time PCR.