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Cloning and expression of pab gene of M. tuberculosis isolated from pulmonary TB patient in E.coli DH5α
"Latar belakang: Antigen38 Mycobacterium tuberculosis merupakan agen serodiagnostik yang potensial karena mengandung
dua epitop spesifik untuk sel B. Mahalnya agen diagnostik menyebabkan lambatnya realisasi diagnosis TB secara cepat di
negara berkembang. Kami memproduksi antigen 38 rekombinan yang berasal dari galur lokal yang kemungkinan dapat
digunakan untuk memproduksi alat serodiagnostik TB yang ekonomis.
Metode: Gen pab diisolasi dari pasien TB paru di Malang, diklon ke plasmid pGEM-Teasy menjadi pMB38. Klon E.coli
DH5α yang membawa pMB38 diseleksi di medium yang ditambah dengan X-Gal. Ekspresi pab dilakukan menggunakan
pMBhis yang berasal dari pPRoExHTc dibawah kontrol promoter Trc dengan inang E.coli DH5α
Hasil: Pencocokan sekuen gen pab dari klon E.coli DH5α berwarna putih dengan gen pab dari M. tuberculosisH37Rv
memperlihatkan homologi sebesar 98%. Protein rekombinan yang sudah dihilangkan signal peptidanya ditemukan di
sitoplasma.
Kesimpulan: Gen pab dari pasien TB dapat diekspresikan secara intraseluler dengan sistem heterolog
Abstract
Background: Mycobacterium tuberculosis antigen38 is a potent serodiagnostic agent containing two M. tuberculosisspecific
B-cell epitopes. The high price of imported diagnostic agents hinders realization of fast clinical TB diagnosis in
developing countries. Therefore, we produced recombinant antigen38 (recAg38M) from M. tuberculosis local strain, which
might be used to produce economical tuberculosis serodiagnostic kit.
Methods: Pab gene that was isolated from pulmonary TB patient in Malang was cloned into a plasmid vector (pGEMTeasy)
to construct pMB38. The E.coli DH5α clone carrying pMb38 was selected on X-gal medium. The expression of pab
was mediated using pPRoExHTc under the control of Trc promoter and E.coli DH5α as host.
Results: Alignment of the pab sequence from the white E.coli DH5α clones with that of M. tuberculosis H37Rv showed
98% homology. The recombinant protein in which the signal peptide has been deleted to prevent the protein being secreted
into medium was found in the cytoplasm.
Conclusion: pab gene of M. tuberculosis isolated from a TB patient could be expressed in heterologous system in E.
coliDH5α."
[Fakultas Kedokteran Universitas Indonesia, Universitas Brawijaya. Fakultas Matematika dan Ilmu Pengetahuan Alam], 2011
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Artikel Jurnal  Universitas Indonesia Library
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Desi Indria Rini
Pengklonaan dan ekspresi gen mce1A (Rv0169) mycobacterium tuberculosis strain beijing dan H37Rv pada escherichia coli BL21 untuk pengembangan kandidat vaksin tuberkulosis = Cloning and expression of mce1A gene of mycobacterium tuberculosis beijing and h37rv strain in expression system escherichia coli bl21 for tuberculosis vaccine candidate development
"Tuberkulosis sejak lama merupakan salah satu penyebab utama kematian manusia karena penyakit infeksi, terutama didaerah yang dilanda kemiskinan dan malnutrisi. Penyakit ini menyerang banyak organ pada tubuh manusia terutamanya adalah paru-paru. Peningkatan jumlah kasus tuberkulosis dipengaruhi oleh infeksi HIV dan resistensi terhadap berbagai macam kombinasi obat. Di Indonesia, infeksi M. tuberculosis oleh strain Beijing diyakini memiliki penyebaran yang paling luas dibandingkan dengan strain lainnya. BCG merupakan vaksin tunggal yang digunakan untuk pencegahan tuberkulosis, namun daya proteksi dan efikasinya berbeda-beda. Protein Mce1A merupakan protein yang diduga berperan penting pada hal invasi dan pertahanan M. tuberculosis didalam makrofag. Beberapa studi telah melakukan penelitian ini, namun di Indonesia belum pernah dilakukan penelitian mengenai ekspresi protein Mce1A Mycobacterium tuberculosis strain Beijing sebagai isolat lokal. Oleh karena itu, pada penelitian ini akan dilakukan pengklonaan dan ekspresi protein Mce1A Mycobacterium tuberculosis strain Beijing lokal dan strain standar H37Rv sebagai pembanding. Gen Mce1A M. tuberculosis strain Beijing dan H37Rv diamplifikasi dengan teknik PCR dan diinsersikan kedalam vektor pET28a. Escherichia coli BL21 kemudian ditransformasi dengan plasmid rekombinan tersebut. Protein Mce1A rekombinan diekspresikan dengan induksi IPTG. E. coli BL21 berhasil ditransformasi dengan plasmid rekombinan yang mengandung sisipan gen Mce1A dengan arah orientasi dan kerangka baca yang benar. Tidak ada mutasi yang ditemukan pada asam amino yang menjadi epitope pengenalan sel B dan sel T. Hasil ekspresi protein Mce1A pada E.coli BL21 menunjukkan pita protein yang lebih tinggi dari seharusnya. Konfirmasi keberadaan protein dilakukan menggunakan teknik Western Blot dengan anti-his detector. Protein Mce1A rekombinan yang telah berhasil diekspresikan pada E.coli BL21 diduga berada dalam bentuk dimer. Hal ini dapat digunakan sebagai data awal kondisi ekspresi untuk pengembangan vaksin subunit pada penelitian berikutnya.
For past centuries until nowadays, tuberculosis remains the leading cause of death in the world from infectious disease wherever poverty, malnutrition and poor housing prevail. Tuberculosis is primarily a disease of the lungs, but may spread to other sites or proceed to a generalized infection. The wide spread of tuberculosis has been further aggravated by another infection disease such as HIV-AIDS and drug resistance. Many strain of Mycobacterium tuberculosis caused tuberculosis infection in Indonesia, but Beijing strain are the most. Bacille Calmette-Guerin (BCG) is the current vaccine for tuberculosis but it has different protection function and efficacy. According to function analysis, mce1A gene predicted has a role in host invasion by Mycobacterium tuberculosis and survival of the pathogen in human macrophages. Several studies abroad have done this research, but in Indonesia, study about protein expression of Mce1A gene of Mycobacterium tuberculosis Beijing strain as local isolates has not much being done. Therefore, in this study we will performed cloning and protein expression of Mce1A gene Mycobacterium tuberculosis Beijing strain as local isolate and standard strain H37Rv as a comparison on expression vector Escherichia coli BL21. Mce1A gene from M. tuberculosis Beijing and H37Rv strain was amplified by PCR and inserted in the vector pET28a. E. coli BL21 then transformed with the recombinant plasmid. Mce1A recombinant protein then expressed with IPTG induction. This study indicate that E. coli BL21 succesfully transformed with a recombinant plasmid containing the Mce1A gene insertion with correct orientation and reading frame. There is no mutation found in the amino acids sequence for B and T cell epitope. Mce1A expression in E. coli BL21 showed protein bands that higher than expected. The protein was confirmed with western blotting using anti-his detector. We assume that Mce1A recombinant protein that have been expressed in E. coli BL21 is in dimeric form. This explanation should be valuable in further studies of expression at the protein level and exposure of proteins on the cell surface of M. tuberculosis under different experimental conditions."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2013
T59195
UI - Tesis Membership  Universitas Indonesia Library
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Pasaribu, Munawaroh
Perbandingan cross priming amplification CPA dan PCR konvensional laboratorium mikrobiologi klinik FKUI untuk deteksi M tuberculosis pada sputum pasien tersangka TB paru tanpa dan dengan HIV/AIDS = Comparison of cross priming amplification CPA and conventional PCR of clinical microbiology FKUI in detection of M tuberculosis in sputum of suspected pulmonary TB patient non HIV/AIDS and HIV/AIDS
"Latar belakang: Diagnosis tuberkulosis (TB), khususnya pada pasien HIV masih merupakan masalah tersendiri, terutama pada daerah dengan sumber daya terbatas. Pemeriksaan mikroskopis hapusan bakteri tahan asam (BTA) merupakan metode yang sederhana dan cepat tetapi hanya mendeteksi 30% -40% kasus Tb sedangkan kultur (baku emas) membutuhkan waktu pemeriksaan yang berminggu-minggu. Metode genotipe (PCR dan isothermal amplification) memiliki sensitivitas yang tinggi dan kerjanya cepat tetapi metode ini masih sangat kompleks dan membutuhkan peralatan khusus. Cross-priming amplification (CPA) merupakan metode amplifikasi DNA secara isothermal dengan menngunakan multiprimer dan enzim polymerase dengan tehnik pembacaan hasil amplifikasi yang sederhana.
Tujuan: Penelitian ini bertujuan untuk mengetahui perbedaanCPA dan PCR TB LMK FKUI dalam mendeteksi M. tuberculosis pada sputum pasien tersangka TB tanpa/dengan HIV.
Metode: 20 sputum pasien non-HIV tersangka TB dan 37 sputum pasien HIV tersangka TB diperiksa dengan CPA dan PCR TB LMK FKUI.
Hasil: Semua yang terdeteksi positif dengan CPA juga dideteksi positif oleh PCR tetapi 20% hasil yang terdeteksi negatif oleh CPA terdeteksi sebagai positif di PCR dan semua yang terdeteksi negatif oleh PCR terdeteksi negatif juga di CPA sedangkan hasil negatif di CPA (20%) masih terdeteksi positif oleh PCR. Dalam mendeteksi M. tuberculosis pada sputum pasien HIV/AIDS tersangka TB paru Terdapat hubungan bermakna antara CPA dengan PCR TB LMK FKUI, hampir semua hasil positif oleh CPA (94.1%) juga dideteksi positif oleh PCR tetapi 30% hasil negatif oleh CPA terdekteksi sebagai positif oleh PCR dan hampir semua hasil negatif oleh PCR (93.3%) terdeteksi negatif juga oleh CPA.
Kesimpulan: Terdapat hubungan bermakna antara CPA dan PCR TB LMK FKUI dalam mendeteksi M. tuberculosis pada sputum pasien tersangka TB paru. PCR TB LMK FKUI lebih sensitif dibanding CPA dalam mendeteksi M.tuberculosis.
Background: The diagnosis tuberculosis (TB) especialy in HIV patients remains a major obstacle to global control of TB, especially in resource limited settings. Light microscopy in sputum smears as common method in detection acid-fast bacilli (AFB) is specific but it only detects 30% to 40% of TB patients, while culture methods as gold standart in TB diagnostic require several weeks of incubation time. Genotypic method (polymerase chain reaction (PCR) and isothermal amplification) is known very sensitive and works fast but it requires special equipment and complex protocols in amplifying and detection amplified products. Cross-priming amplification (CPA) principle is isothermal amplification using multiple cross-linked primers (six to eight primers) and detection of amplified products is performed on special design plastic which is easy to performed and identified.
Objective: The study aimed to determine difference of PCR and CPA to detect M. tuberculosis in sputum specimens from suspected pulmonary tuberculosis without/with HIV patients.
Methods: 20 sputum samples suspected pulmonary TB of non-HIV patients and 37 sputum samples suspected pulmonary TB of HIV patients were subjected to CPA and PCR TB LMK FKUI.
Results:. All samples which were positive by CPA were also PCR positive but 20% result that were CPA negative were still positive by PCR and all samples with PCR negative were negative detected by CPA while some samples that were negative by CPA were still positive detected by PCR (20%). In HIV/AIDS population, there were significant correlation between CPA and PCR TB LMK FKUI which all positive result of CPA (94.1%) were also PCR positive but 30% of CPA negative were still CPA positive and almost all of PCR negative (93.3%) were CPA negative.
Conclusion: There were significant correlation between CPA and PCR TB LMK FKUI in ditected of M.tuberculosis in sputum of suspected pulmonary TB in populations of non HIV/AIDS patients and HIV/AIDS patients."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2014
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UI - Tesis Membership  Universitas Indonesia Library
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Ardath Herland S.
Peran metode TB-LAMP dalam mendeteksi mycobacterium tuberculosis dari sputum pasien terduga tuberkulosis paru = The Role of TB-LAMP method in detecting mycobacterium tuberculosis from sputum patients suspected of pulmonary tuberculosis
"Insiden tuberkulosis (TB) di Indonesia merupakan salah satu yang tertinggi di dunia. Penegakan diagnosis secara tepat merupakan salah satu upaya untuk mengendalikan TB. Modalitas uji diagnostik laboratorium TB yang tersedia yaitu pemeriksaan mikroskopis Basil Tahan Asam (BTA) dan pemeriksaan biakan memiliki beberapa keterbatasan. Secara global, terjadi peningkatan dalam penggunaan Tes Cepat Molekuler (TCM) sebagai uji diagnostik laboratorium TB. Salah satu TCM yang telah direkomendasikan oleh World Health Organization (WHO) yaitu loop-mediated isothermal amplification (LAMP). Oleh karena itu, perlu dilakukan penelitian untuk mengevaluasi metode LAMP, sehingga metode tersebut dapat diterapkan secara rutin di Indonesia. Penelitian dilaksanakan di Laboratorium Mikrobiologi Klinik Fakultas Kedokteran Universitas Indonesia (LMK-FKUI) terhadap 100 orang pasien terduga TB paru. Setiap pasien menyerahkan dua sputum langsung, yaitu sputum sewaktu dan sputum pagi/sewaktu. Terhadap setiap sputum langsung kemudian dilakukan pemeriksaan mikroskopis BTA. Dua sediaan sputum langsung dari setiap pasien kemudian digabung untuk menghasilkan mixed sputum. Terhadap setiap mixed sputum dilakukan pemeriksaan mikroskopis BTA, biakan Lowenstein-Jensen (LJ) dan pemeriksaan TB-LAMP. Biakan yang tumbuh diidentifikasi menggunakan tes MPT64. Hasil penelitian menunjukkan nilai kapa (κ) pemeriksaan mikroskopis BTA antara sputum langsung dan mixed sputum sebesar 0,88; p < 0,001 (95% CI 0,78-0,97). Persentase hasil LAMP (+) pada sputum langsung dengan hasil BTA (-) sebesar 28,07%, sedangkan persentase hasil LAMP (+) pada mixed sputum dengan hasil BTA (-) sebesar 32,78%. Metode TB-LAMP memiliki nilai sensitivitas sebesar 100% (95% CI 89,56-100%) dan spesifisitas sebesar 69,64% (95% CI 55,74-80,84%). Nilai duga positif TB-LAMP sebesar 71,19% (95% CI 57,73-81,86%), sedangkan nilai duga negatif TB-LAMP sebesar 100% (95% CI 88,83-100%). Pada hasil mikroskopis BTA yang sesuai (concordant) dengan biakan TB, nilai sensitivitas dan spesifisitas TB-LAMP berturut-turut sebesar 100% (95% CI 89,09-100%) dan 73,58% (95% CI 59,42-84,32). Adapun nilai sensitivitas TB-LAMP pada hasil mikroskopis BTA yang tidak sesuai (discordant) dengan biakan TB, yaitu sebesar 100% (95% CI 19,79-100%). Metode TB-LAMP memiliki nilai sensitivitas dan nilai duga negatif yang tinggi. Untuk menegakkan diagnosis TB paru secara tepat, metode TB-LAMP harus dikombinasikan dengan gejala dan tanda yang terdapat pada pasien.
The incidence of tuberculosis (TB) in Indonesia is one of the highest in the world. Appropriate diagnosis is an effort to control TB. Existing TB laboratory diagnostic test modalities, which are Acid-Fast Bacilli (AFB) smear and culture examination have several limitations. Globally, there has been an increase in the use of the molecular rapid test as a TB laboratory diagnostic test. One of the molecular rapid tests recommended by the World Health Organization (WHO) is loop-mediated isothermal amplification (LAMP). Therefore, research is needed to evaluate the LAMP method, so that the method can be applied routinely in Indonesia. The study was conducted at the Clinical Microbiology Laboratory of the Faculty of Medicine, Universitas Indonesia for 100 patients suspected of pulmonary TB. Each patient handed over two direct sputums, which are the spot sputum and morning/spot sputum. Against each direct sputum, an AFB smear was carried out. Two direct sputum preparations from each patient were then combined to produce mixed sputum. For each mixed sputum, AFB smear, Lowenstein-Jensen (LJ) culture and TB-LAMP examination were carried out. Cultures that were grown were identified using MPT64 tests. The results of the study showed that the kappa (κ) value of AFB smear between direct sputum and mixed sputum was 0.88; p < 0.001 (95% CI 0.78-0.97). The percentage of LAMP (+) in direct sputum with AFB (-) was 28.07%, while the percentage of LAMP (+) in mixed sputum with AFB (-) was 32.78%. The TB-LAMP method had a sensitivity value of 100% (95% CI 89.56-100%) and a specificity of 69.64% (95% CI 55.74-80.84%). Positive predictive value of TB-LAMP was 71.19% (95% CI 57.73-81.86%), while the negative predictive value of TB-LAMP was 100% (95% CI 88.83-100%). In concordant AFB smear results with TB culture, the TB-LAMP sensitivity and specificity values were 100% (95% CI 89.09-100%) and 73.58% (95% CI 59.42-84,32), respectively. The sensitivity value of TB-LAMP on AFB smear results which were discordant with TB culture was 100% (95% CI 19.79-100%). The TB-LAMP method had a high sensitivity value and negative predictive value. To properly diagnose pulmonary TB, the TB-LAMP method must be combined with the symptoms and signs that the patient has."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2019
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UI - Tugas Akhir  Universitas Indonesia Library
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Mimien Harianti
Molecular cloning of the vitellogenin gene in the hard-lipped barb (osteochillus hasseltii c.v) and photoperiod’S effects on gene expression
"Photoperiod affects fish reproduction as it regulates activities of the endocrine glands, which produce the hormones needed for geonadal growth and development, gametogenesis, and reproductive cycles. This study aimed to determine the effects of photoperiod on the hard-lipped barb’s reproductive performance by exposing the fish to three photoperiod treatments (light hour: L, darks hour: D), namely 14L:10D (control), 8L:16D (short photoperiod) and 18L:6D (long photoperiod), with four aquaria, each containing 9 fish, serving as replicates. The fish were kept under these photoperiods for 8 weeks. Liver activity, the observable variable in the study, was evaluated by measuring vitellogenin gene expression. Normalized data were then subjected to ANOVA, followed by Tukey’s range test. The hard-lipped barb’s vitellogenin cDNA was found have a 1136 bp sequence and the vitellogenin precursors encoded cDNA comprising 378 amino acids. The vitellogenin gene in each experimental group saw a significant increase on average when exposed to longer photoperiods (P<0.05), and the highest levels of vitellogenin gene expression occurred under long photoperiods (LP, 18 h light:6 h dark). These results indicate that longer photoperiods stimulate and improve the hard-lipped barb’s reproductive performance."
Bogor: Seameo Biotrop, 2018
634.6 BIO 25:3 (2018)
Artikel Jurnal  Universitas Indonesia Library
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Kresanti Dewi Ngadimin
Pengklonaan fragmen DNA gen PD-1 N-terminal pada sistem ekspresi TOP10 E. coli = Cloning of DNA fragment encoding N-terminal region of PD-1 gene from E. coli Top10 expression system
"Protein PD-1 biasanya diekspresikan berlebihan pada pasien kanker dan menghambat sistem imun. Antibodi monoklonal dari PD-1 dapat digunakan untuk menghambat jaras tersebut. Namun, urutan nukelotida dari epitop dengan afinitas yang kuat masih belum diketahui. Oleh karena itu, plasmid yang mengandung epitop kecil dari PD-1 dibuat untuk proses epitope mapping. Tujuan penelitian ini adalah untuk mendapatkan plasmid yang mengandung daerah N-terminal dari gen PD-1. DNA insert sintetik dibuat dari metode PCR dan dipurifikasi. Kedua DNA insert dan plasmid pQE80L didigesti dengan enzim restriksi BamH1 dan HindIII, dipurifikasi dan diligasi. Plasmid tersebut dimasukkan kedalam sel kompeten TOP10 dengan transformasi heat shock. Koloni positif diseleksi menggunakan metode PCR koloni dan verifikasi dengan menggunakan digesti dan sanger sequencing. Produk PCR dari EP1PD1 didapat dengan menggunakan suhu annealing sebesar 57ºC dan berhasil diligasi ke plasmid pQE80L and ditransformasikan. Hasil dari sequencing menunjukan urutan yang sama namun terdapat insersi diawal. Beberapa modifikasi perlu dilakukan untuk mengekspresi protein EP1PD1. Konklusi dari penelitian ini adalah plasmid yang mengandung epitop 1 PD-1 berhasil diperoleh dengan insersi.
PD-1 protein tends to be overexpressed in cancer patient and inhibits immune system. Monoclonal antibody of PD-1 can be used to inhibit this pathway. However, the sequence of epitope with strong affinity is currently unknown. Thus, plasmid containing small epitope of PD-1 was made for PD-1 epitope mapping process. The objective of this research is to obtain plasmid containing N-terminal region of PD-1 gene. Synthetic insert DNA was made using PCR method and purified. Both insert DNA and pQE80L plasmid were digested using BamH1 and HindIII enzyme, purified and ligated. It was then inserted into TOP10 competent cell using heat shock transformation method. Positive colonies are selected using PCR colony and verified using digestion and Sanger sequencing. PCR product of EP1PD1 are obtained using annealing temperature of 57ºC and able to be ligated to pQE80L plasmid and transformed. Sequencing result shows EP1PD1 result with insertion in the beginning. Modifications are required to express EP1PD1 protein. In conclusion, the plasmid containing Epitope 1 of PD-1 are able to be obtained with insertion."
Depok: Fakultas Kedokteran Universitas Indonesia, 2019
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UI - Skripsi Membership  Universitas Indonesia Library
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Reny Guspratiwi
Perbadingan kloning dan dan ekspresi gen antigen tuberkulosis 85B dengan menggunakan signal peptide original dan signal peptide AQ1 endoxilanase = Comparison of cloning and expression of tuberculosis antigen 85B gene using signal peptide original and signal peptide AQ1 endoxylanase
"Ekspresi gen antigen 85B perlu diketahui sebelum dijadikan kandidat vaksin tuberkulosis yang diharapkan bisa menggantikan vaksin BCG. Ekspresi tersebut bisa diketahui setelah gen antigen 85B diklona dan ditransformasikan ke dalam E. coli DH5α. Ekspresi gen diharapkan bisa lebih banyak dengan penggunaan signal peptide AQ1 endoxilanase. Gen antigen 85B yang berukuran 991 pb diklona ke dalam vektor pUC57 fragmen 2 yang berukuran sekitar 2500 pb, sehingga menghasilkan plasmid rekombinan yang berukuran sekitar 3500 pb. Hasil ekspresi dari plasmid rekombinan diuji dengan SDS-PAGE. Hasil penelitian menunjukkan gen antigen 85B berhasil diklona ke dalam vektor pUC57 fragmen 2, namun antigen tersebut belum berhasil diekspresikan.
The expression of antigen 85B gene needed to be known before the antigen is used as tuberculosis vaccine candidate that is expected to replace the BCG vaccine. The expression of antigen 85B gene could be known after the gene was cloned and transformed into E. coli DH5α. The expression of the gene was expected to increase in number with the use of AQ1 endoxylanase signal peptide. Antigen 85B gene size 991 bp was cloned into a vector pUC57 fragmen 2 with size approximately 2500 bp, resulting in a recombinant plasmid with size approximately 3500 bp. The expression results of recombinant plasmid were tested with SDS-PAGE. The results showed antigen 85B gene successfully cloned into the vector pUC57 fragment 2, but these antigens have not been successfully expressed."
Depok: Universitas Indonesia, 2014
S55800
UI - Skripsi Membership  Universitas Indonesia Library
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Fadhilah Yuza Fitri
Korelasi Klinis dan Hasil Uji Kepekaan Jamur Aspergillus yang Diisolasi dari Sputum Pasien Tuberkulosis Paru = Correlation of Clinical Characteristics and Susceptibility Study of Aspergillus Isolated from Sputum of Pulmonary Tuberculosis Patients
"Latar belakang: Pasien tuberkulosis (TB) paru rentan mengalami infeksi oportunistik, termasuk oleh Aspergillus (aspergilosis paru). Keberadaan Aspergillus dikonfirmasi dengan uji kultur. Uji kepekaan Aspergillus terhadap obat anti-jamur (OAJ) dilakukan untuk mengetahui pilihan OAJ yang tepat. Itrakonazol merupakan salah satu OAJ pilihan untuk aspergilosis paru. Resistensi OAJ dapat disertai gejala klinis yang luas. Penelitian ini bertujuan untuk mengetahui karakteristik klinis pasien dikaitkan dengan hasil uji kepekaan Aspergillus terhadap itrakonazol.
Metode: Penelitian potong-lintang ini dilakukan pada Juli-November 2021 dan merupakan bagian dari penelitian sebelumnya. Isolasi Aspergillus dari sputum pasien TB paru dilakukan menggunakan medium agar Sabouraud. Karakteristik klinis yang diteliti meliputi usia, jenis kelamin, indeks massa tubuh (IMT), dan gejala. Adapun uji kepekaan jamur dilakukan dengan metode cakram sesuai protokol.
Hasil: Penelitian ini menyertakan 28 isolat Aspergillus sp. yang berasal dari sputum 28 pasien TB paru. Jumlah pasien laki-laki lebih dominan (24 orang). Rerata usia 50 ± 15,1 tahun, dengan kelompok usia terbanyak < 60 tahun (21 pasien). Sebanyak 12 pasien (42,9%) memiliki IMT rendah. Gejala klinis yang didapatkan meliputi: batuk (42,9%), batuk darah (35,7%), sesak (39,3%), nyeri dada (14,3%), dan rasa lelah (35,7%). Kultur sputum menunjukkan pertumbuhan 28 isolat Aspergillus, terdiri atas: 14 isolat Aspergillus fumigatus, 6 isolat Aspergillus flavus, dan 8 isolat Aspergillus niger. Uji kepekaan Aspergillus terhadap itrakonazol menunjukkan 23 isolat sensitif, 3 isolat intermediat, dan 2 isolat resisten. Analisis statistik menunjukkan tidak terdapat hubungan bermakna antara karakteristik klinis pasien dengan hasil uji kepekaan jamur.
Kesimpulan: Tidak terdapat hubungan bermakna antara karakteristik klinis pasien dengan hasil uji kepekaan Aspergillus terhadap itrakonazol dalam penelitian ini.
Introduction: Pulmonary tuberculosis (TB) patients are susceptible to opportunistic infections, including Aspergillus infections (aspergillosis). The presence of Aspergillus was confirmed by a culture test, followed by its susceptibility study to antifungal. Antifungal resistance is generally accompanied by serious symptoms, so clinical observations are important for better clinical awareness. This study aims to determine the relationship between clinical characteristics and susceptibility study of Aspergillus to itraconazole.
Method: This cross-sectional study was carried out from July-November 2021, as part of the previous study on aspergillosis in TB patients. Aspergillus was isolated from the sputum of pulmonary TB patients using Saboraud's agar dextrose medium. Clinical characteristics obtained through patient’s history including age, gender, body mass index, and symptoms The fungal susceptibility test was carried out by disc diffusion method according to the protocol after treatment.
Result: This study included 28 isolates of Aspergillus from the sputum of 28 pulmonary TB patients. The number of males were dominant (24 from 28 patients) than females. The mean age was 50 ± 15.1 years, with the commonest age group < 60 years (21 patients). Total of 12 patients (42.9%) had a low body mass index. Clinical symptoms included: cough (42.9%), hemoptysis (35.7%), dyspnea (39.3%), chest pain (14.3%), and fatigue (35.7%). %). The sputum culture showed 14 Aspergillus fumigatus isolates, 6 Aspergillus flavus isolates, and 8 Aspergillus niger. The susceptibility test of Aspergillus to itraconazole revealed 23 sensitive isolates, 3 intermediate isolates, and 2 resistant isolates. Statistical analysis showed that there was no relationship between the patient’s clinical characteristics and the antifungal susceptibility test of itraconazole.
Conclusion: There was no significant correlation between the patient’s clinical characteristics and the antifungal susceptibility test to itraconazole in this study"
Depok: Fakultas Kedokteran Universitas Indonesia, 2022
S-pdf
UI - Skripsi Membership  Universitas Indonesia Library
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Titi Sekarindah
Pengaruh pemberian Vitamin A terhadap profil imun pasien Tuberkulosis Paru dengan terapi obat anti Tuberkulosis = The effect of vitamin A supplementation on the immune profile of pulmonary Tb patient during oral anti tuberculosis treatment
"ABSTRAK
Ruang Lingkup dan Cara Penelitian : Penyakit tuberkulosis paru masih merupakan masalah di negara berkembang termasuk Indonesia. Tuberkulosis menduduki urutan ke 2 sebagai penyebab kematian menurut hasil survey nasional 1992. Dari kepustakaan diketahui bahwa pada penderita tuberkulosis didapati kelainan imunitas seluler, sehingga untuk penyembuhan penyakit tuberkulosis diperlukan pengaktifan sistem imun testa imunitas seluler. Vitamin A sudah lama dikenal sebagai imunomodulator. Dari penelitian terdahulu pemberian retinoid dapat meningkatkan respon imun seluler antara lain kenaikan sel T penolong dan T penolong/supresor. Pada penelitian ini diharapkan pemberian vitamin A sejumlah 2x 200000IU pada penderita TB paru dengan OAT dapat meningkatkan imunitas seluler. Tujuan penelitian ini adalah menilai pengaruh pemberian vitamin A pada penderita tuberkulosis paru yang sedang mendapat OAT terhadap jumlah limfosit total, limfosit T total, sub populasi limfosit T, kadar retinol plasma, dan keadaan klink penderita. Vitamin A 200.000 IU diberikan pada awal penelitian dan setelah 4 minggu. Penelitan dilakukan secara uji klinik tersamar ganda pada 40 penderita TB paru. Penderita dibagi dalam 2 kelompok masing-masing 20 orang yang diberi vitamin A dan placebo. Pada akhir penelitian yaitu setelah 8 minggu, ada 5 orang drop out.
Hasil dan kesimpulan : Dari 40 orang peserta penelitian 10% kadar retinol plasma rendah (<20pg/dl), 30%normal, rendah(20-30pg/d.l), 60% normal. Pada pemeriksaan imunitas seluler 53,85% ada gangguan dan 46,15% normal. Nilai rata rata hitung (X) retinal plasma kelompok placebo dan perlakuan sebelum pemberian vit. .A/placebo berturut-turut adalah 30,24 ± 7,51 µg/dl dan 30,82 ±7,31 µg/dl. Setelah pemberian adalah 36,85 ± 9,74 µg/dl dan 38,02 ± 8,29 µg/dl. Pada uji t berpasangan dari kelompok perbkkan kenaikannya bermakna (p
ABSTRACT
Scope and Method of Study : Pulmonary tuberculosis is still a major health problem in the developing countries including Indonesia. Tuberculosis is number 2 as cause of death (National Survey's data, 1992). According to literature study tuberculosis patients are suffering from an immune defect. To recover from the disease the immune response especially the cellular immune response needs to be activated, because mycobacterium TB are living intracellular. Vitamin A is known as an immunomodulator. From earlier research it is known that retinoid could enhance cellular immune response, ie. increasing T helper cells and the ratio Thelperffsupresor. The hypothesis is that supplementation of vitamin A 2x2000001U to pulmonary TB patients could increase the cellular immunity. The aim of this study was to asses the vitamin A supplementation on the immune?s profile of pulmonary TB patient who are on oral anti tuberculosis treatment. Plasma retinot, nutrients intake, BMI, clinical findings were examined. Vitamin A 200.000M was given twice, in the beginning of the study and after 4 weeks. The design of the study was a randomized double blind clinical trial. Forty patients were selected and divided into 2 groups, a placebo and treatment (vitamin A) group. At the end of the study (after the 8th week), 5 patients dropped out.
Findings and Conclusions : Among 40 patients 10% showed plasma ret noK20 p g/dl), 30% normal low (20-30pgldl) and 60% normal. (03011g041). The cellular immunity was 53,85% abnormal and 46,15% normal The means (X) of plasma retinol of the placebo and study group before supplementation were 30.24 ± 7,51 µg/dl and 30.82 ± 7.31µg/dl respectively; after supplementation 36.85±9.74µg/dl and 38.02 ± 8.29µgldl respectively. Statistical analysis using paired t test showed that the study group was increasing s' 0,05), however there was no Significant difference between the 2 groups. The mean (X) of total lymphocyte before supplementation of the placebo and study group were 22.61 ± 6.51% and 22.63 ± 8,62%; after supplementation 38.09 ± 19.91% and 35.20 + 10.71%. Both were increasing significant; however there was no significant difference between the 2 groups. The T lymphocyte, T helper and ratio Thelper CT supresor were decreasing. T helper more in the placebo group 5.75% 2.29% but there was no significant difference. This study concluded that although vitamin A supplementation 2 X 200.000 IU could increase the plasma retinol but could not yet improve the immune response and clinical status significantly."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 1996
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Bimo Satrio Putra Erlyandi
Kloning dan Ekspresi Gen Envelope 2 (E2) Virus Chikungunya (CHIKV) Isolat Indonesia pada Saccharomyces cerevisiae INVSc1 = Cloning and Expression of Envelope 2 (E2) Gene Chikungunya Virus Isolate from Indonesia in Saccharomyces cerevisiae INVSc1
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Manifestasi klinis seseorang yang terinfeksi virus chikungunya (CHIKV) mirip dengan seseorang yang terinfeksi virus dengue (DENV) sehingga menyulitkan para praktisi kesehatan dalam melakukan diagnosis penyakit. Badan Pengkajian dan Penerapan Teknologi (BPPT) telah memulai pengembangan kit diagnostik infeksi CHIKV berbasis protein rekombinan untuk mendeteksi keberadaan partikel CHIKV dalam serum darah. Penelitian sebelumnya, BPPT telah berhasil memproduksi serum antibodi poliklonal anti-CHIKV sebagai langkah awal pengembangan kit diagnostik. Penelitian kloning dan ekspresi gen envelope 2 (E2) virus chikungunya (CHIKV) Isolat Indonesia pada Saccharomyces cerevisiae merupakan penelitian lanjutan dengan tujuan untuk memperoleh plasmid rekombinan pYES2-E2 CHIKV dan untuk memperoleh protein rekombinan E2 yang diekspresikan oleh Saccharomyces cerevisiae. Protein rekombinan E2 yang dihasilkan akan diuji reaktivitasnya dengan menggunakan antibodi poliklonal anti-CHIKV yang telah diproduksi BPPT sebelumnya. Hasil penelitian menunjukkan bahwa plasmid rekombinan pYES2-E2 CHIKV telah berhasil diperoleh berdasarkan 2 metode verifikasi yaitu metode PCR dan metode digesti. Hasil analisis ekspresi protein dengan SDS PAGE menunjukkan pita protein pada berbagai macam ukuran. Analisis protein dengan western blotting memberikan hasil dengan munculnya pita protein pada kisaran ukuran 25--35 kDa. Protein rekombinan E2 CHIKV diduga telah berhasil diperoleh dan memerlukan pengujian lebih lanjut dengan menggunakan antibodi poliklonal anti-CHIKV.

Diagnosis of people-infected chikungunya virus (CHIKV) is quite challenging since it has similar symptoms with dengue virus infection. Badan Pengkajian dan Penerapan Teknologi (BPPT) had started the development of diagnostic kit chikungunya virus infection based on recombinant protein to improve the diagnosis of CHIKV infection. Previous research, BPPT had successfully produced policlonal antibody anti-CHIKV serum. Cloning and expression of envelope 2 (E2) gene chikungunya virus isolate from Indonesia in Saccharomyces cerevisiae is aimed to produce recombinant plasmid pYES2-E2 CHIKV and to produce recombinant protein E2 expressed by S. cerevisiae.  The results were recombinant plasmid pYES2-E2 CHIKV had successfully achieved based on 2 verification methods (PCR and digestion method). Protein expression analysis by western blotting gave result a single band appearance suspected E2 CHIKV protein with molecule size ranged from 25 to 35 kDa. Sample which positively contains recombinant protein E2 CHIKV is continued to test its reactivity with policlonal antibody anti-CHIKV serum which had previously produced by BPPT.

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Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2020
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