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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 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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ABSTRACTSyphilis 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."

"Sifilis 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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Syphilis 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."

"Latar Belakang: Permasalahan gigi dan mulut pada anak di usia mixed dentition di Indonesia masih tinggi, salah satunya di daerah Jawa Barat. Permasalahan gigi dan mulut dapat disebabkan oleh status kesehatan oral yang buruk. Status kesehatan oral dapat dinilai dari nilai OHI-S. Status kesehatan mulut buruk ditandai dengan penumpukan plak (salah satunya plak supragingiva) dapat menyebabkan peningkatan jumlah bakteri, salah satunya bakteri Veillonella parvula. Bakteri Veillonella parvula berperan dalam proses adhesi dan metabolisme bakteri late colonizer seperti Treponema denticola yang berperan dalam patogenesis penyakit periodontal. Bakteri Veillonella parvula dan Treponema denticola memiliki protein berupa VtaA dan Msp yang berperan dalam proses adhesi ke biofilm. Sampai saat ini, belum ada penelitian yang meneliti hubungan kedua protein tersebut dengan status kebersihan rongga mulut. Tujuan: Mengetahui apakah ada hubungan dan korelasi antara ekspresi gen Msp dan VtaA dengan status kebersihan rongga mulut. Metode: Penelitian menggunakan 40 sampel plak supragingiva yang diambil dari anak berusia 9-12 tahun di SD Sukaluyu dan dikelompokkan berdasarkan kategori skor OHI-S. Sampel kemudian diekstraksi RNA dan DNA, lalu dianalisis menggunakan Real Time PCR. Hasil kuantifikasi DNA dianalisis menggunakan absolute quantification untuk mengidentifikasi jumlah bakteri, sedangkan hasil kuantifikasi RNA dianalisis menggunakan relative quantification untuk membandingkan ekspresi gen. Hasil: Terdapat korelasi positif bermakna (p=0.008) antara jumlah bakteri Treponema denticoladengan memburuknya nilai OHI-S. Terdapat perbedaan bermakna antara jumlah bakteri Treponema denticola pada OHI-S sedang dan buruk (p=0.016). Korelasi positif tidak signifikan ada pada jumlah bakteri Veillonella parvula dan ekspresi gen Msp, sedangkan korelasi negatif tidak signifikan ditemukan pada VtaA. Kesimpulan: Ada hubungan antara menambahnya jumlah bakteri Treponema denticola seiring dengan memburuknya status kesehatan oral. Tidak ditemukan korelasi dan perbedaan antara jumlah bakteri Veillonella parvula, ekspresi gen Msp dan VtaA jika dibandingkan dengan kategori OHI-S.

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Latar Belakang: Dental and oral problems in children with mixed dentition in Indonesia are still highly prevalent, especially in West Java. These problems are caused by poor oral health status, which can be assessed by OHI-S score. Poor oral health status, which is characterized by plaque accumulation, can cause the quantity of bacteria in mouth to increase. One of those bacterias is Veillonella parvula, a bacteria which plays a role in the adhesion process and metabolism of late colonizer bacteria. Treponema denticola is a late colonizer bacteria which contributes to the progression of periodontal diseases. In order to adhere to a biofilm, Treponema denticola produces protein called Msp, while Veillonella parvula produces VtaA. However, the relationship between these proteins to oral health status has not been well studied. Objective : The aim of this study is to analyze the relationship and correlation between Msp and VtaA gene expression and oral health status. Methods: 40 samples are collected from supragingival plaque of children between the ages of 9-12 years old in SD Sukaluyu and grouped into 3 categories (poor, moderate, good) based on each samples’s OHI-S score. Samples are then extracted and analized by real-time PCR. DNA quantification results are analyzed using absolute quantification to identify the amount of bacteria present. RNA quantification results are analyzed using relative quantification to identify each gene expression relative to calibrator samples. Results: There is a significant positive correlation (p=0.008) found between the quantity of Treponema denticola and OHI-S score. A significant difference (p=0.016) is found between the amount of Treponema denticola in moderate OHI-S and poor OHI-S category. There is a non-significant positive correlation between the amount of Veillonella parvula and Msp gene expression and OHI-S score. VtaA gene expression showed a non-significant negative correlation. Conclusion: This study demonstrated there is a relationship between the increasing quantity of Treponema denticola and the worsening state of oral health status. There is no relationship between Msp and VtaA gene expression and the quantity of Veillonella parvula and oral health status."