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"Latar belakang :Berdasarkan data Laboratorium Mikrobiologi Klinik FKUI tahun 2009, P. aeruginosa dan A. baumanii yang resisten terhadap beberapa golongan antibiotik terutama karbapenem merupakan patogen nosokomial terbanyak di ICU RSUPNCM. Penyusunan kebijakan penggunaan antibiotik dan pengendalian infeksi bakteri MDR memerlukan data tentang mekanisme resisten yang banyak terjadi pada kedua isolat tersebut.Tujuan Umum: Mengetahui karakteristik fenotip dan genotip P.aeruginosa dan A.baumanii resisten karbapenem yang diisolasi dari pasien ICU RSUPN Cipto Mangunkusumo tahun 2011.Metode: Penelitian ini merupakan penelitian pendahuluan yang bersifat deskriptif retrospektif. Sampel adalah isolat stok P. aeruginosa dan A. baumanii , yang menunjukkan hasil uji kepekaan rutin intermediet atau resisten terhadap satu atau lebih antibiotik golongan karbapenem. Setelah isolat dihidupkan, dilakukan identifikasi ulang dengan uji biokimia konvensional dan dilakukan uji konfirmasi penghasil karbapenemase dengan metode Modifikasi Hodge dan deteksi gen pengkode dihasilkannya enzim karbapenemase yaitu blaKPC-2; blaIMP-1; blaVIM-2; blaNDM-1dan blaOXA-48 menggunakan metode PCR.Hasil: Terdapat isolat stok P. aeruginosa sejumlah 77 dan A. baumanii 85. Berdasarkan hasil identifikasi ulang didapatkan 20 isolat P.aeruginosa dan 42 isolat A.baumanii yang resisten terhadap karbapenem. Hasil uji fenotif penghasil karbapenemase positif pada 4 isolat P. aeruginosa dan 16 isolat A. baumanii. Deteksi gen pengkode dihasilkannya enzim karbapenemase menunjukkan bahwa terdapat 1 isolat (5%) P. aeruginosa memiliki gen blaKPC-2, 4 isolat (20%) memiliki gen blaIMP-1; 1 isolat (5%) memiliki blaVIM-2 dan 2 isolat(10%) memiliki blaNDM-1. Pada isolat A. baumanii ditemukan blaKPC-2 dan blaVIM-2 masing masing pada 1 isolat (5%). Sementara itu gen resisten blaOXA-48 tidak ditemukan pada kedua spesies bakteri.Kesimpulan: Pada isolat P. aeruginosa dan A. baumanii resisten karbapenem yang diisolasi dari pasien ICU-RSUPN Cipto Mangunkusumo tahun 2011 didapatkan isolat penghasil enzim karbapenemase dengan gen pengkode blaKPC-2;blaVIM-2;blaIMP-1 dan blaNDM-1.

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Background : Based on data from Laboratory of Clinical Microbiology, Faculty of Medicine Universitas Indonesia in 2009, P. aeruginosa and A. baumannii which were resistant to multiple classes of antibiotics, especially to carbapenem, were the most prevalent nosocomial pathogens in ICU RSUPNCM. The policies formulation of controlling antibiotic usage and MDROs infection requires data on the mechanism of resistance in both isolates.

Aim : To explain the phenotype and genotype characteristics of carbapenem resistant P.aeruginosa and A.baumanii isolated from ICU patients, RSUPN Cipto Mangunkusomo in 2011.

Method : This is a preliminary descriptive retrospective study. Samples were laboratory isolated stock of P. aeruginosa and A. baumannii, which were intermediate or resistant to one or more classes of carbapenem antibiotics in routine antibiotic susceptibility test. After re-inoculating the isolates, re-identification were done by conventional biochemical testing, then confirmation test conducted by Modified Hodge test and detection of karbapenemase produced encoding genes, blaKPC-2; blaIMP-1; blaVIM-2;blaNDM-1 and blaOXA-48, using PCR method.

Result : Of 77 isolates of P. aeruginosa isolates and 85 isolates of A. baumannii, 20 isolates P. aeruginosa and 42 isolates A. baumanii were resistant to carbapenem. By carbapenemase producing phenotypic test, positive results showed in 4 isolates of P. aeruginosa and 16 isolate A. baumannii. The detection of karbapenemase produced encoding genes showed that there was 1 isolate (5%) P. aeruginosa had blaKPC-2 gene, 4 isolates (20%) had blaIMP-1 gene; 1 isolates (5%) had blaVIM-2 gene and 2 isolates (10%) had blaNDM-1 gene. In A. baumannii population found blaKPC-2 gene and blaVIM-2 gene at 1 isolates (5%), respectively. Meanwhile, blaOXA- 48 gene were not found in both species of bacteria

Conclusion: In isolates of P. aeruginosa and A. baumannii carbapenem-resistant from in the ICU Cipto Mangunkusomo in 2011 producing isolates obtained with the gene encoding the enzyme karbapenemase blaKPC-2; blaVIM-2; blaIMP-1 and blaNDM-1."

"Bakteri yang terdapat dalam usus manusia berada dalam keseimbangan dan memainkan peranan penting dalam fungsi metabolisme dan imunologi tubuh, Infeksi yang terjadi pada saluran cerna, seperti diare, dapat mengakibatkan terjadinya ketidakseimbangan pada komposisi bakteri usus tersebut. Pengetahuan mengenai profil mikroba usus pada kasus diare anak usia tertentu memiliki manfaat yang penting dalam memberikan informasi awal untuk pengembangan tata laksana kasus diare yang berkaitan dengan pengembalian keseimbangan mikroba usus. Penelitian ini merupakan penelitian deskriptif analitik dengan disain potong lintang. Feses dikumpulkan dari dua kelompok subyek penelitian, dengan diare dan tanpa diare dari anak-anak usia 2-12 tahun di Jakarta Utara. Sampel kemudian di ekstraksi dengan kit QIAmp® DNA Stool Mini untuk kemudian dilakukan deteksi dan identifikasi bakteri dengan menggunakan polymerase chain reaction / Electrospray Ionization-Mass Spectrometry. Secara keseluruhan diperoleh 80 subjek, terdiri dari 33 anak-anak yang mengalami diare (subyek diare) dan 47 anak-anak yang tidak mengalami diare (subjek non-diare). Tiga puluh dari 33 sampel dalam kelompok diare terdeteksi keberadaan bakteri. Enam dari 33 sampel memberikan hasil multiple matches, sedangkan 3 sampel lainnya tidak terdeteksi adanya bakteri. Pada kelompok nondiare, di 28 dari 47 sampel terdeteksi adanya bakteri, hasil multiple matches pada 8 dari 47 sampel dan 13 sampel tidak terdeteksi adanya bakteri. Dalam kedua kelompok didominasi oleh Echerechia coli dan juga diikuti oleh Klebsiella pneumonia. Keragaman bakteri yang terdeteksi pada kelompok diare (12 dari 30 sampel) lebih dari pada kelompok non-diare (5 dari 28). Filum bakteri yang dideteksi pada kelompok sampel diare adalah Firmicutes (5 sampel), Proteobacteria (24), Bacteroidetes (1), dan di kelompok non diare adalah Actinobacteria (2), Proteobacteria (25), Verrucomicrobia (1). Hubungan antara enteropatogen dengan kejadian diare tidak signifikan secara statistik (p= 0,571, uji Chi-square), akan tetapi terdapat hubungan yang kuat antara risiko kejadian diare yang disebabkan oleh enteropatogen (OR = 0,724 dengan 95% CI: 0,237-2,215). Hasil penelitian menyimpulkan bahwa keragaman bakteri yang dideteksi pada kelompok diare lebih dari pada kelompok non-diare dengan adanya kesamaan dalam pola bakteri yang paling banyak terdeteksi pada kedua kelompok sampel, adanya temuan bakteri anggota filum Actinobacteria (Bifidobacterium longum) yang bersifat probiotik pada kelompok non diare dan tampaknya kemungkinan anak-anak yang positif enteropatogen pada fesesnya memiliki kecenderungan untuk mengalami diare dibandingkan dengan yang tidak.

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Microbiota present in the human intestinal are diverse and play important roles in metabolism and immunology. Infection that occurs in gastrointestinal tract, may lead to an imbalance in the composition of the intestinal bacteria. Knowledge on the intestinal microbes profile in children at spesific age with and without diarrhea might shed a light in the management of diarrhea associated with intestinal microflora imbalance. The objective this study is to obtain a profile of intestinal bacteria in children at spesific age with diarrhea and non-diarrhea which may be important for initial information in management of diarrhea associated intestinal microbes imbalance.

This study was an analitical descriptive with cross sectional design. Stool samples were collected from two groups of subjects, with diarrhea and without diarrhea in children of 2-12 years old in North Jakarta. The samples were extracted using QIAamp® DNA Stool Mini Kit first followed by detection and identification using Polymerase Chain Reaction / Electrospray Ionization-Mass Spectrometry. A total 80 subjects were obtained, consisted of 33 children with diarrhea (diarrhea subjects) and 47 children without diarrhea (non-diarrheal subjects). Thirty of the 33 stool samples in diarrhea group showed the presence of one species microorganism (complete match), 6 samples resulted in multiple matches, while the other three samples did no show any bacteria.

In the non-diarrhea group, of total 47 stool samples, 28 showed the presence of single match bacteria, 8 specimens gave result of multiple matches and 13 specimens showed no detectable bacteria. In both groups Echerechia coliand Klebsiella pneumonia appeared to be dominant. The bacteria present in the diarrhea group (12 of 30 samples) were more diverse than in nondiarrheal group (5 of 28). Phyla found in diarrhea group consisted of Firmicutes (5 samples), Proteobacteria (24), Bacteroidetes (1), while in non-diarrhea group were Actinobacteria (2), Proteobacteria (25), Verrucomicrobia (1).

The conclusion is bacteria detected in diarrhea group apparently were more diverse than in nondiarrhea. There was similarity in the pattern of most detected bacteria in both sample groups, however, member of Actinobacteria (Bifidobacterium longum) where detected only in non-diarrhea group. Likely the chance of children with enteropathogen detected in the stool would have diarrhea more than children with no enteropathogen detected."

"ABSTRAK Latar belakang : Cytomegalovirus (CMV) merupakan salah satu infeksi oportunistikpada pasien dengan sindrom immunodefisiensi (AIDS). Gejala klinis dan CT scantidak dapat menegakkan diagnosa definitif ensefalitis CMV. Oleh karena itudiperlukan uji alternatif untuk menegakkan diagnosis infeksi CMV pada pasien HIVdengan infeksi otak. Salah satu uji yang sensitif dan spesifik adalah Real TimePolymerase Chain Reaction (rPCR).Tujuan : Mendapatkan uji deteksi molekular CMV pada pasien HIV dengantersangka infeksi otak.Metode : Penelitian dilakukan dalam 3 tahap. Tahap 1 adalah optimasi konsentrasiprimer, probe, suhu annealing, volume elusi ekstraksi DNA, dan volume cetakan.Tahap 2 adalah uji spesifisitas (reaksi silang) dan uji sensitivitas (ambang batasdeteksi DNA) rPCR dan tahap 3 adalah penerapan uji rPCR yang sudah dioptimasiterhadap sampel plasma, urin, dan LCS.Hasil : Kondisi optimal uji rPCR telah diperoleh dengan konsentrasi primer danprobe 0,1 μM, dengan kondisi suhu reaksi rPCR: aktivasi enzim pada 950C selama 3menit; 45 siklus pada 950C selama 15 detik (denaturasi) dan 560C selama 1 menit(annealing dan ekstensi). Volume elusi ekstraksi DNA yang optimal untuk ketigajenis sampel (LCS, plasma dan urin) adalah 40 μL, dan volume cetakan rPCR untukLCS, plasma, dan urin, masing-masing adalah 5, 4, dan 3 μL. Uji rPCR mampumendeteksi DNA pada 50.000 jumlah kopi/mL dan tidak bereaksi silang denganStaphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacteriumtuberculosis, Candida spp, Toxoplasma gondii, EBV,HSV,dan VZV. Penerapan ujirPCR pada sampel klinis memberikan hasil negatif pada semua sampel LCS, 72,22%positif pada sampel plasma, dan 72,22% positif pada sampel urin.Kesimpulan: Telah dilakukan optimasi uji rPCR dengan minimal deteksi DNACMV 50.000 jumlah kopi/mL dan tidak bereaksi silang dengan mikroorganisme yangberpotensi menyebabkan positif palsu (false positive).ABSTRACT Background: Cytomegalovirus (CMV) is one of opportunistic infections in patientswith Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are nottypical, and CT scans can not define encephalitis CMV specifically. Therefore, it isimportant to apply an alternative assay for sensitive and specific detection of CMVinfection in HIV patients with suspected central nervous system (CNS) infections.One of the assays is real time polymerase chain reaction (rPCR).Objective: To obtain a molecular assay for detection of CMV in HIV patients withsuspect CNS infections.Methods: This study was conducted in three phases. The first is optimization ofconcentrations of primers, probe, annealing temperature, final elution of DNAextraction, and volume of PCR template. The second is determinations of sensitivity(minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR,and the third is application of the rPCR for clinical samples of plasma, urine, andliquor cerebrospinal (LCS).Results: The rPCR reaction showed optimal concentrations of primers and probe at0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing andextension). Final elution of DNA extraction was 40 μL and volume of PCR templatesfor urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimaldetection of DNA at 50,000 copies/mL and was not cross-reacted withStaphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacteriumtuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), HerpesSimplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR forclinical samples showed that the rPCR yielded 72.22% positive for plasma or urine,and negative for all LCS samples.Conclusion: The rPCR has been optimized in this study with minimal DNA detectionat 50,000 copies/mL and was not cross-reacted with other microorganisms that arepotential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patientswith Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are nottypical, and CT scans can not define encephalitis CMV specifically. Therefore, it isimportant to apply an alternative assay for sensitive and specific detection of CMVinfection in HIV patients with suspected central nervous system (CNS) infections.One of the assays is real time polymerase chain reaction (rPCR).Objective: To obtain a molecular assay for detection of CMV in HIV patients withsuspect CNS infections.Methods: This study was conducted in three phases. The first is optimization ofconcentrations of primers, probe, annealing temperature, final elution of DNAextraction, and volume of PCR template. The second is determinations of sensitivity(minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR,and the third is application of the rPCR for clinical samples of plasma, urine, andliquor cerebrospinal (LCS).Results: The rPCR reaction showed optimal concentrations of primers and probe at0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing andextension). Final elution of DNA extraction was 40 μL and volume of PCR templatesfor urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimaldetection of DNA at 50,000 copies/mL and was not cross-reacted withStaphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacteriumtuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), HerpesSimplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR forclinical samples showed that the rPCR yielded 72.22% positive for plasma or urine,and negative for all LCS samples.Conclusion: The rPCR has been optimized in this study with minimal DNA detectionat 50,000 copies/mL and was not cross-reacted with other microorganisms that arepotential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patientswith Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are nottypical, and CT scans can not define encephalitis CMV specifically. Therefore, it isimportant to apply an alternative assay for sensitive and specific detection of CMVinfection in HIV patients with suspected central nervous system (CNS) infections.One of the assays is real time polymerase chain reaction (rPCR).Objective: To obtain a molecular assay for detection of CMV in HIV patients withsuspect CNS infections.Methods: This study was conducted in three phases. The first is optimization ofconcentrations of primers, probe, annealing temperature, final elution of DNAextraction, and volume of PCR template. The second is determinations of sensitivity(minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR,and the third is application of the rPCR for clinical samples of plasma, urine, andliquor cerebrospinal (LCS).Results: The rPCR reaction showed optimal concentrations of primers and probe at0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing andextension). Final elution of DNA extraction was 40 μL and volume of PCR templatesfor urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimaldetection of DNA at 50,000 copies/mL and was not cross-reacted withStaphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacteriumtuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), HerpesSimplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR forclinical samples showed that the rPCR yielded 72.22% positive for plasma or urine,and negative for all LCS samples.Conclusion: The rPCR has been optimized in this study with minimal DNA detectionat 50,000 copies/mL and was not cross-reacted with other microorganisms that arepotential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patientswith Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are nottypical, and CT scans can not define encephalitis CMV specifically. Therefore, it isimportant to apply an alternative assay for sensitive and specific detection of CMVinfection in HIV patients with suspected central nervous system (CNS) infections.One of the assays is real time polymerase chain reaction (rPCR).Objective: To obtain a molecular assay for detection of CMV in HIV patients withsuspect CNS infections.Methods: This study was conducted in three phases. The first is optimization ofconcentrations of primers, probe, annealing temperature, final elution of DNAextraction, and volume of PCR template. The second is determinations of sensitivity(minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR,and the third is application of the rPCR for clinical samples of plasma, urine, andliquor cerebrospinal (LCS).Results: The rPCR reaction showed optimal concentrations of primers and probe at0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing andextension). Final elution of DNA extraction was 40 μL and volume of PCR templatesfor urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimaldetection of DNA at 50,000 copies/mL and was not cross-reacted withStaphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus,Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacteriumtuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), HerpesSimplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR forclinical samples showed that the rPCR yielded 72.22% positive for plasma or urine,and negative for all LCS samples.Conclusion: The rPCR has been optimized in this study with minimal DNA detectionat 50,000 copies/mL and was not cross-reacted with other microorganisms that arepotential to cause false positive results."

"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."

"Uveitis adalah inflamasi intraokular yang terjadi pada saluran uvea mata ataupun jaringan yang berada di dekatnya, diantaranya retina atau vitreous. Uveitis merupakan penyebab kebutaan nomor tiga di dunia dengan prevalensi tertinggi pada kelompok umur pekerja aktif 20-50 tahun . Selama ini diagnosis uveitis infeksi dan non-infeksi di Indonesia ditegakkan secara klinis dan didukung pemeriksaan serologi darah. Pemeriksaan antibodi pada cairan akuos dan serum serta marka genetik, khususnya HLA B-27 belum pernah dilakukan. Penelitian ini dilakukan untuk memberikan alternatif pendukung diagnosis klinis yaitu pemeriksaan deteksi molekuler HLA B-27 dan deteksi antibodi dengan perhitungan koefisien Goldmann-Witmer. Subjek penelitian adalah pasien uveitis aktif di Poliklinik Infeksi dan Imunologi RSCM Kirana yang berjumlah 79 orang. Berdasarkan koefisien Goldmann-Witmer, mikroba penyebab uveitis adalah Toxoplasma gondii dan virus Varicella zoster. Gen HLA-B27 ditemukan pada enam subjek penelitian dan terdapat kesesuaian dengan kasus uveitis non-infeksi tetapi belum dapat dibuktikan kemaknaannya secara statistik.

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Uveitis is an intraocular inflammation which occurs on the uvea or its surrounding tissue such as retina or vitreous. Uveitis is known as the third major cause of blindness in the world with the highest prevalence is productive age group 20 50 years old . To date, infectious and non infectious uveitis stipulation in Indonesia is based on clinical examination and serology test. The aqueous and serum antibody titer measurement as well as genetic marker examination, especially on HLA B27, has never been done before. In this research we perform HLA B27 detection through molecular analysis and antibody titer measurement to know the Goldmann Whitmer coefficient. Sample size used in this research is 79 patients with active uveitis obtained from ldquo Poliklinik Infeksi dan Imunologi RSCM Kirana rdquo . The analysis of Goldmann Whitmer coefficient showed that the pathogens responsible for uveitis are Toxoplasma gondii and Varicella zoster. HLA B27 were found in six patients and there is similiarity between non infectious uveitis but this value does not have prove statistically. "

"Leptospirosis adalah penyakit infeksi akut yang dapat menyerang rnanusia maupun hewan yang disebabkan bakteri Leprospfra spp dan digolongkan sebagai zoonosis. Gejala klinis leptospirosis yang tidak spesiiik dan sulitnya uji laboratorium untuk konfirmasi diagnosis mengakibatkan penyakit ini seringkali tidak terdiagnosis. Oleh karena itu dalam ponelirian ini dilakukan optimasi uji diagnostik molekuler menggimakan real-time PCR sebagai deteksi cepat, sensitif dan spesiflk untuk Leptospira patogen pada manusia DNA bakten di dalam spesimen darah diekstraksi menggunak:an QIAamp DNA Blood Mini Kit, Qiagen dan spesimen urin diekstraksi menggunakan QIAamp DNA Stool Mini Kit,Qiagen dengan prosodur sesuai dengan petunjuk manualnya. Primer dan probe yang digunakan berdasarkan publikasi penelitian oleh Smythe dkk, 2002. Dari hasil uji optimasi kondisi optimal real-time PCR didapat suhu annealing 60°c, konsentrasi primer 0,9 uM dan konsentrmi probe 0,2 uM. Spesifisitas primer diuji menggunakan DNA balcteri patogen lain Hasii uji sensitiiitas real-time PCR untuk mendeteksi konsentrasi DNA terendah bakteri Leprospim spp adalah 0,75 fypl, hasil uji spesitisitas real-time PCR menunjukkan bahwa primer yang digunakan untuk deteksi balderi Leprospira spp tidak beraksi silang dengan genom bakteri-bakteri uji, konsentrasi minimal DNA bakteri yang masih terdeteksi dalam darah mencapai 150 fg/pl, sedangkan dalam urin mencapai 1470 fg/pl yang masih dapat dideteksi dengan pemeriksaan real-time PCR. Metode real-time PCR ini dapat digunakan sebagai alternatif pemeriksaan mikrobiologi yang cepat dan tepat untuk mendiagnosis leptospirosis.

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Leptospirosis is an emerging infectious disease in human and animals caused by Leptospira spp. and considered endemic in Indonesia due to its tropical climate. The International Leptospirosis Society (2001) declared Indonesia has high incidence of leptospirosis and ranked the third in the world for mortality (16.7%) The clinical features are not specific and may result in a missed or delayed diagnosis. The microbiology diagnostic method e.g. culture and microscopic agglutination test (MAT) are sensitive and specific but time-consuming and high cost. The other method to detect the antibody result false positive reactions and need confirmation by the MAT. Therefore in this study we optimized the real-time PCR assay, which has been used to detect a large number of microbes. It has high sensitivity and specificity, thus making it ideal as a rapid and accurate method to detect pathogen Leptospira spp. in human specimens. The amplification of the DNA control was performed optimally with the following conditions: annealing temperature is 60°C, primer volume is 0.5p1 (final concentration: 0.9 phd); probe volume is 0.2 ul (final concentration 0.2 pM). This method may detect the DNA in the Mastermix Mix with the concentration of 0.75 fg/ul, however in blood specimen the limit of detection of the DNA 150 fg/pl and in urine is 1470 fg/pl. The primer used in this assay is not complementary with the DNA of other pathogenic Leptospira spp. The real-time PCR assay is a rapid and accurate method to detect pathogenic Leptospira in human specimens. Further studies are needed to know the sensitivity and specificity of the real-time PCR assay compared to other diagnostic methods in clinical settings."

"Legionellosis is a collection of infection that emerged in the second half of the 2Oth century, and that are caused by Legionella pneumophila and related bacteria. Legionellosis consists of two clinical syndromes, Legionnaires?disease is characterisized by pneumonia and pontiac fever is self-limiting, influenza like illness. Outb According this study, the sensitivity of duplex PCR to detect Legionella pneumophila in sterile NaCl 0.9% is 2.8 CFU/ml. The sensitivity of the duplex PCR in seeded water samples are 62 CFU/400ml of tap water sample, 32 CFU/400ml of sterile distiiled water and 32 CFU/400 ml of sterile NaCl 0.9%. The culture method in this study can not recovered Legionella from seeded water samples. The presence of Legionella .spp and Legionella pneumophila in cooling tower water was investigated using the duplex PCR. Of 9 cooling tower water sample and 3 tap water sample, 8 were positive for Legionella spp, 1 were positive for Legionella pneumophila and 3 were negative. According detection Legionella in seeded water samples and cooling tower water, the culture method can not be used to recover Legionella, but the duplex PCR can be used as rapid detection for Legionella spp and Legionella pneumophila.

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Legionella pneumophila merupakan penyebab utama lgionellosis yang mulai muncul pada pertengahan abad 20. Legionellosis dapat berkembang menjadi dua keadaan klinik, pertama Legionnares? disease yang merupakan penyakit multi sistem pneumonia, kedua Pontiac fever suatu penyakit mirip dengan flu dan dapat sembuh dengan sendirinya. Umumnya kasus legionellosis terjadi akibat dari kontaminasi pada sistem air panas maupun dingin pada gedung bertingkat seperti cooling tower, kondensor, spa, kolam renang, Oleh karena itu deteksi bakteri Legionellla pada sistem air di gedung bertingkat dan rumah sakit diperlukan untuk mencegah legionellosis nosokomial ataupun komunitas. Deteksi legionella dengan metode konvensional memerlukan media khusus dan waktu inkubasi yang lama. Pada penelitian ini duplex PCR dikembangkan untuk mendeteksi Legionella spp dan Legionella pneumophila pada sampel air cooling lower, dengan primer dari sekuens gen 16S rRNA unluk mendeteksi Legionella spp Serta primer sekuens gen nano untuk mendeteksi Legionella pneumophila. Pada penelitian ini Duplex PCR dapat digunakan untuk mendeteksi Legionella pneumophila dalam suspensi NaCl 0.9% hingga batas deteksi 2,8 CFU/ml. Hasil uji simulasi menggunakan sampel air yang ditambahkan pengenceran berseri Legionella pneumophila menunjukkan batas deteksi hingga 62 CFU/ 400 ml air kran, 32 CFU/400 ml akuadest steril dan 32 CFU/ 400 ml NaCl 0.9% stril. Hasil uji sirnulasi dengan metode kultur tidak menunjukkan pertumbuhan koloni pada agar BCYE plus. Hasil uji coba Duplex PCR terhadap 9 sampel air cooling tower dan 3 sampel air kran adalah satu sampel menunjukkan pita spesiiik L. pneumophia, 8 sampel yang menunjukkan pita spesifik Legionella spp dan 3 sampel negatif. Berdasarkan uji simulasi dan pemeriksaan sampel air cooling lower; metode kultur pada penelitian ini belum dapat mendeteksi keberadaan bakteri Legionella, sedangkan deteksi Legionella spp dan L. pneumophila dapat dilakukan dengan metode duplex PCR."

"Munculnya varian virus influenza yang berpotensi menimbulkan pandemi merupakan kejadian yang tidak terduga dan jarang terjadi. Penanganan dalam situasi seperti ini membutuhkan produksi vaksin skala besar dalam waktu singkat seperti pada kasus virus influenza H1N1 pdm09. Salah satu strategi yang dapat dilakukan adalah produksi vaksin subunit dengan memanfaatkan sistem ekspresi ragi untuk memproduksi protein antigen rekombinan. Tujuan penelitian ini adalah mengekspresi protein fusi HA1-MA2-NS1 rekombinan untuk pengembangan vaksin subunit. Plasmid pPICZαA-HA1-MA2-NS1 linear diintegrasikan ke dalam genom Pischia pastoris strain GS115 melalui rekombinasi homolog. Proses transformasi ini dilakukan dengan metode elektroporasi dan menghasilkan 13 klon ragi yang mengandung multiple integran gen fusi HA1-MA2-NS1. Analisis bioinformatika menduga protein fusi memiliki berat molekul 75-85 kDa. Visualisasi protein pada supernatan dan pelet dengan SDS-PAGE memperlihatkan adanya pita protein dengan ukuran yang diharapkan. Hasil western blot pada pellet mendeteksi ukuran protein target antara 42-135 kDa. Pemurnian protein fusi dengan NI-NTA gagal dilakukan dan diduga karena sedikitnya jumlah protein fusi yang terbentuk sehingga tidak terdeteksi pada visualisasi SDS-PAGE.

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The emergence of influenza virus variants that could potentially cause a pandemic is an unforeseen occurrence and rare. Handling a situation like this requires a large-scale vaccine production in a short time as in the case of the H1N1 virus pdm09. One strategy that can be done is a subunit vaccine production by utilizing a yeast expression system to produce recombinant protein antigens.

The purpose of this study was to express a recombinant proteins of fusion HA1- MA2-NS1 for subunit vaccine development. PPICZαA HA1-MA2-NS1 linear plasmid integrated into the genome Pichia pastoris strain GS115 through homologous recombination. This transformation process is carried out by electroporation method and generating 13 yeast clones containing multiple genes HA1-MA2-NS1 integrant. Bioinformatics analysis of protein suspect fusion protein has a molecular weight of 75-85 kDa. Visualization of proteins in the supernatant and pellet by SDS-PAGE showed protein bands with sizes expected.

But western blot results in the pellet detect the target protein size between 42-135 kDa. Purification of fusion proteins with NI-NTA were failed due to small amount of fusion protein that undetected on SDS-PAGE visualization."