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"ABSTRAKSifilis merupakan penyakit multistadium kronik yang disebabkan oleh bakteriTreponema pallidum dan ditularkan dari lesi aktif pasangan seksual atau dari ibuhamil yang terinfeksi pada janin yang dikandungnya. Saat ini telah terjadipeningkatan kasus T. pallidum resisten azitromisin akibat mutasi titik A2058Gdan A2059G pada gen 23S rRNA. Di Indonesia belum ada data terkait resistensiT. pallidum terhadap azitromisin sehingga penelitian ini bertujuan untukmendapatkan metode nested multipleks PCR untuk deteksi kedua mutasi yangmenyebabkan resistensi. Tiga pasang primer digunakan pada reaksi nested PCR.Untuk mendapatkan kondisi uji yang optimal dilakukan optimasi parameter yangpenting pada proses PCR. Uji nested multipleks PCR dapat mendeteksi 22.000jumlah copy DNA/ml dan tidak bereaksi silang terhadap mikroorganisme yangpotensial menyebabkan hasil positif palsu. Uji awal 45 sampel klinis darahditemukan 13 sampel positif T. pallidum dan tidak ditemukan mutasi baikA2058G maupun A2059G. Dua sampel positif dikonfirmasi dengan DNAsekuensing dan menunjukkan tidak ada mutasi titik. Uji nested multipleks PCRyang telah dikembangkan pada penelitian ini dapat digunakan untuk deteksimutasi gen 23S rRNA T. pallidum yang menyebabkan resistensi azitromisin pada sampel klinis darah.

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ABSTRACTSyphilis is a chronic, multi-stage infectious disease caused by Treponemapallidum that is usually transmitted sexually by contact with an active lesion of a partner or congenitally from an infected pregnant woman to her fetus.Azithromycin-resistant strains of T. pallidum is associated with a single pointmutation (either A2058G or A2059G) in both copies of the 23S rRNA gene of T.pallidum. These strains are now prevalent in many countries but there is no dataavailable about it in Indonesia. Therefore, in this study we developed a nestedmultiplex PCR to detect A2058G and A2059G 23S rRNA gene point mutations ofT. pallidum. Three primer sets were designed for nested PCR reactions. To obtainoptimal PCR reaction, all parameters were optimized. The assay could detect atleast 22000 DNA copy number/ml and showed no cross reaction with othermicroorganisms that potentially cause false positive result. A total 13 of 45 wholeblood specimens were PCR positive for T. pallidum and no single point mutation(either A2058G or A2059G) were detected by PCR. Two positive specimens wereconfirmed by DNA sequencing and showed no mutation. Thus, nested multiplexPCR developed in this study is potential to detect azithromycin-resistant T.pallidum in whole blood samples."

"ABSTRAKSifilis merupakan penyakit multistadium yang ditularkan terutama melalui hubungan seksual. Saat ini penggunaan uji polymerase chain reaction (PCR) untuk Treponema pallidum telah banyak digunakan dan diharapkan mampu mengurangi masalah dalam uji diagnostik sifilis. Hasil uji PCR Treponema pallidum dipengaruhi oleh jenis spesimen, metode PCR dan gen target. Penelitian ini ditujukan untuk menilai penggunaan darah dan serum untuk uji multiplex nested PCR dengan gen target 23S rRNA Treponema pallidum. Studi potong lintang dilakukan dari bulan April 2015 - April 2016. Pengambilan sampel secara konsekutif dari pasien dengan gambaran klinis sifilis sekunder yang datang ke poliklinik Infeksi Menular Seksual (IMS) di Jakarta. Uji PCR dilakukan terhadap 122 spesimen klinis (61 darah dan 61 serum). Uji serologi rapid plasma reagin (RPR) dan Treponema pallidum Haemagglutination Assay (TPHA) dilakukan pada semua serum. Hasil positif uji PCR darah sebesar 22,95% dan serum sebesar 6,56%, sedangkan hasil positif uji serologi sebesar 68,85%. Pada hasil uji serologi positif, proporsi hasil positif uji multiplex nested PCR Treponema pallidum darah sebesar 30,95% dibandingkan serum 9,52%. Uji PCR terhadap darah mampu mendeteksi 3,25 kali lebih tinggi daripada serum. Penggunaan darah memberikan nilai kepositivan yang lebih tinggi dibandingkan serum pada uji multiplex nested PCR Treponema pallidum menggunakan gen target 23S rRNA

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ABSTRACTSyphilis is a multistage disease transmitted primarily through sexual intercourse. Nowadays, polymerase chain reaction (PCR) test for Treponema pallidum has been widely used and expected to overcome problems in diagnostic test for syphilis. The PCR Treponema pallidum are influenced by type of specimens, PCR methods and gene targets. This study is aim to assess the use of blood and serum using multiplex nested PCR Treponema pallidum targeting 23S rRNA. Cross-sectional study was conducted from April 2015 - April 2016. Sampling was carried out consecutively from patients with clinical features of secondary syphilis who came to sexual transmitted infection (STI) clinics in Jakarta. PCR test performed on 122 clinical specimen ( 61 blood and 61 serum). All serum were tested with RPR and TPHA assay. The positive results of PCR test on blood was 22,95% and serum was 6,56%, while the positive results of serology was 68,85%. On positive serological test results, the proportion of positive results of multiplex nested PCR Treponema pallidum on blood was 30,95% compared to serum 9,52%. PCR test on blood is able to detect 3,25 times higher than serum. The use of blood give a higher positivity compared to serum in multiplex nested PCR Treponema pallidum using 23S rRNA gene target."

"Sifilis adalah penyakit menular seksual kronik yang memiliki manifestasi klinis yang bervariasi dan menetap untuk waktu yang lama. Neurosifilis merupakan salah satu komplikasi sifilis sistemik dengan temuan di cairan serebrospinal dengan atau tanpa gejala yang jelas. Pemeriksaan yang saat ini tersedia dalam mendukung diagnosis hanya tersedia pemeriksaan analisis cairan serebrospinal dan serologi Treponema pallidum. Saat ini belum diketahui prevalensi neurosifilis di rumah sakit peneliti dan profil serologi Treponema pallidum dari bahan cairan serebrospinal. Penelitian ini merupakan penelitian potong lintang, dilakukan November 2017-Maret 2018 terhadap 50 cairan serebrospinal dan darah yang diperiksakan analisis cairan serebrospinal dengan keterangan klinis terduga infeksi intrakranial. Serum dan cairan serebrospinal diperiksakan RPR, TPHA, anti-Treponema pallidum ELISA IgG dan khusus cairan serebrospinal diperiksa pula rapid test Treponema pallidum. Uji statistik menggunakan chi quare and Fisher exact test.Dari penelitian terhadap 50 cairan serebrospinal dan serum didapatkan rapid test Treponema pallidum, RPR dan TPHA cairan serebrospinal reaktif 4(8%). Dari bahan serum didapatkan RPR reaktif 8(16%) dan TPHA reaktif 9(18%). Anti-Treponema pallidum ELISA IgG positif 4 sampel (8%). Dari 50 sampel didapatkan 7 (14%) neurosifilis, 4 confirmed neurosyphilis dan 3 probable neurosyphilis sesuai kriteria Center for Disease Control and Prevention. Profil analisis cairan serebrospinalnya tidak berwarna, jernih, tidak ada bekuan, hitung sel 12.71 ±9.20 sel/μl, dominasi mononuklear 11.57±9.47 sel/μl, Pandy positif, protein cairan 42.29±21.49 mg/dl, glukosa cairan 55±5.16 mg/dl, glukosa serum 101.04±20.10 mg/dl, dan klorida 122.14±2.48 mEq/L. Pemeriksaan RPR, TPHA, dan anti-Treponema pallidum ELISA IgG dengan bahan serum dan cairan serebrospinal memiliki hubungan bermakna. Dari penelitian ini didapatkan 14% sesuai dengan neurosifilis dari populasi penelitian dan didapatkan 85.71% dengan HIV reaktif. Pada pasien HIV disarankan RPR dan TPHA serum untuk pemeriksaan skrining sifilis.

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Syphilis is a chronic sexually transmitted disease that has varying clinical manifestations and persist for a long time. Neurosyphilis is one of the complications of systemic syphilis with findings in cerebrospinal fluid with or without obvious symptoms. Examinations currently available for diagnostic support were cerebrospinal fluid analysis and serology of Treponema pallidum. There is currently no known prevalence of neurosyphilis in the research hospital and serologic profile of Treponema pallidum from cerebrospinal fluid. This study was a cross sectional study, conducted November 2017-March 2018 against 50 cerebrospinal fluid and blood samples that examined cerebrospinal fluid analysis with clinical information of suspected intracranial infection. Serum and cerebrospinal fluid examined by RPR, TPHA, anti-Treponema pallidum ELISA IgG and particulary rapid test Treponema pallidum for cerebrospinal fluid. Statistic tests were chi quare and Fisher exact test.From a total of 50 cerebrospinal fluid and serum, 4(8%) had reactive cerebrospinal fluid T. pallidum rapid tests, RPRs and TPHAs. From serum there were 8(16%) reactive RPRs and 9(18%) reactive TPHAs. Anti-Treponema pallidum ELISA IgG was positif 4 samples (8%). Among the 50 samples, 7 (14%) had neurosyphilis, 4 were confirmed neurosyphilis and 3 were probable neurosyphilis according to Center for Disease Control and Prevention criteria. The cerebrospinal fluid analysis profile is colorless, clear, without clot, cell count 12.71±9.20 cells/μl, mononuclear 11.57±9.47 cells/μl, positive for Pandy, cerebrospinal fluid protein 42.29±21.49 mg/dl, glucose 55±5.16 mg/dl, serum glucose 101.04±20.10 mg/dl, and chloride 122.14±2.48 mEq/L. Rapid Plasma Reagin, TPHA, and anti-Treponema pallidum ELISA IgG were associated between serum specimen and cerebrospinal fuid. Neurosyphilis was found in 14% of our patient population and 85.71% was reactive for HIV. Rapid Plasma Reagin and TPHA in sera were recommended for syphilis screening for HIV patient."