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ANALISIS PRIMER UNTUK AMPLIFIKASI PROMOTER inhA MULTIDRUG RESISTANCE TUBERCULOSIS (MDR-TB) DENGAN METODE POLYMERASE CHAIN REACTION (PCR) I Gusti Ayu Agung Septiari; Putu Sanna Yustiantara; Sagung Chandra Yowani
Jurnal Kimia (Journal of Chemistry) Vol. 9, no. 1 Januari 2015
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (202.804 KB) | DOI: 10.24843/JCHEM.2015.v09.i01.p19

Abstract

The aim of this study was to analyze several primary combinations for amplifying inhA promoter region by in silico and in vitro ways. Primary in silico’s analysis was done by Clone Manager Suite 6 program. fabG gene sequence of M. tuberculosis was downloaded from www.ncbi.nlm.nih.gov (genbank: U66801.1) and used as DNA template. In vitro detection was done by PCR technique using P16 and 86 M. tuberculosis MDR isolates as DNA template. Amplification was done in described conditions: predenaturation at 95°C for 15 minutes, 45 cycle of amplication (denaturation on 94°C for 1 minute, annealing on 54°C for 1 minute 20 seconds dan extension on 72°C for 1 minute 10 seconds) and also post extension on 72°C for 10 minutes. PCR product was detected by agarose gel elektroforesis (1,5%). In conclusion, combination of primary forward (mabA-inhA-promoter-FS) 5’-ACATACCTGCTGCGCAAT-3’ (18 nucleotide) and primary reverse (mabA-inhA-promoter-R) 5’-CTCCGGTAACCAGGACTGAA-3’ (20 nucleotide) (Chen et al., 2011) have met the good criteria of primary combination which was seen from several aspects such as: primary length, Tm value, %GC, stability, number of hairpins, dimers and runs. In vitro detection showed that the primary combination also amplified inhA promoter region with the length of 284 pb
AMPLIFIKASI FRAGMEN 0,4 KB DAERAH D-LOOP DNA MITOKONDRIA LIMA INDIVIDU SUKU BALI TANPA HUBUNGAN KEKERABATAN DENGAN METODE POLYMERASE CHAIN REACTION (PCR) K. Ratnayani; Sagun Chandra Yowani; Liangky Syane S
Jurnal Kimia (Journal of Chemistry) Vol. 3, No. 1 Januari 2009
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

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Abstract

The human mitochondrial DNA (mtDNA) has higher polimorfism level than nucleous genom, and it ismaternally transmitted. D-Loop is a non-sense region in human mtDNA that has the highest polimorfism. Generally,the aim of this research is to find out the variation in D-Loop region of mtDNA in some Balinese without familycorrelation. For that reason, this research was brought out to determine the sequences of nucleotide of D-Loop regionin five normal Balinese from different families without correlation. The specific aim of this research is to amplify the0,4 kb fragment of mtDNA D-Loop region in five Balinese above, using the PCR methode. In conducting the PCR,the temperature of annealing of primer and the weight of template of mtDNA were optimized. Several phases thathave been conducted : 1). Lisis of the cavum oris epithelium; 2). Quantation of DNA; 3). Reaction PCR; 4). Result ofPCR detection with agarosa gel electroforesisThe result of this research is the amplification of 0,4 kb fragment of D-Loop region in mtDNA by PCR.This research also found the optimum temperature annealing, which was 55 0C, and the optimum weight of templateof DNA which was ± 0,688 ?g.
DESAIN PRIMER UNTUK AMPLIFIKASI GEN katG MULTIDRUG RESISTANCE TUBERCULOSIS (MDR-TB) DENGAN METODE POLYMERASE CHAIN REACTION (PCR) Putu Tedi Suryadi; Ketut Ratnayani; Sagung Chandra Yowani
Jurnal Kimia (Journal of Chemistry) Vol. 8, No. 1 Januari 2014
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (235.693 KB) | DOI: 10.24843/JCHEM.2014.v08.i01.p13

Abstract

Tuberculosis, the world’s major diseases, is one of the emerging infectious disease. The tuberculosis problem has become complicated and burdensome due to the emergence of drug resistant such as isoniazid (INH) resistant strains of Mycobacterium tuberculosis. Mutation in katG gene is the main mechanism of INH-resistance in most strains. Amplification of M. tuberculosis katG gene was performance by using PCR for detect the mutation. A pair of specific PCR primers (forward and reverse) was the most important factor to limit the target region of amplification. Primer designing is preceedly carried out for producing the specific primer desired. The aim of this study was to design the specific primers for a fragment of katG gene. In silico primer design was carried out by using Clone Manager Suite 6. DNA sequence template used in this primer design was downloaded from www.ncbi.nml.nih.gov. M. tuberculosis H37Rv katG gene with genbank code X68081.1 was choosen. This study was successfully designed the forward primer (5' GAAGTACGGCAAGAAGCTCTC 3') and reverse one (5' CGTGATCCGCTCATAGATCG 3') which was length of 21 and 20 nucleotide, respectively. These pair of primers were meet the requirement of a good primer include primer length, Tm value, percentage of GC content, no hairpins, limited dimers and runs. In conclusion, the result of this research showed that the primer designed were acceptable to amplify the fragment of 724 bp of katG gene in silico.
PERBANDINGAN KUALITAS DNA DENGAN MENGGUNAKAN METODE BOOM ORIGINAL DAN BOOM MODIFIKASI PADA ISOLAT Mycobacterium tuberculosis 151 Dewi Andayani Farmawati; I Nengah Wirajana; Sagung Chandra Yowani
Jurnal Kimia (Journal of Chemistry) Vol. 9, no. 1 Januari 2015
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (66.834 KB) | DOI: 10.24843/JCHEM.2015.v09.i01.p07

Abstract

Tuberculosis (TB) is a disease caused by the Mycobacterium tuberculosis. Isolation  DNA is a necessary step to obtain the bacterial chromosomal DNA used in the Polymerase Chain Reaction (PCR). Boom isolation method is an isolation method commonly used in isolation DNA of M. tuberculosis. In Bali, the isolation DNA of M. tuberculosis conducted at the Laboratory of Biomolecular FK UNUD uses boom modification method. This research aims to compare the quality of DNA produced by the Boom methods and Boom modification methods. This research was started with the isolation process using Boom method and Boom modification  and subsequently amplified by PCR. Detection of PCR products was performed with electrophoresis method. DNA quality was determined by the thickness of bands DNA PCR product and purity analysis by spectrophotometry UV-Vis. The results obtained show that the quality of DNA Mycobacterium tuberculosis 151  isolate using Boom modification method (Laboratory of Biomolecular FK UNUD) is relatively better than Boom original methods (Boom et al, 1990).
KARAKTERISASI TOXOPLASMA GONDII ISOLAT INDONESIA Sagung Chandra Yowani; Endang Kumolosasi; Marlia Singgih Wibowo
Jurnal Kimia (Journal of Chemistry) Vol. 1, No. 1 Januari 2007
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

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Abstract

Cathodic protection basically reduces the corrosion rate of a metallic structure by reducing its corrosionpotential, Toxoplasma gondii isolated from diaphragm of sheep at an abbatoir in Sukabumi, West Java had beencharacterized by Centre Research Institute for Animal Sciences. The characterization included study of morphologyby optical microscope, study of ultrastructure by transmission electron microscope, the study of the parasite growthin mice Mus musculus, and study of proteins of the parasite. The growth of parasite in mice had been studied usingtwo groups of mice i.e., normal group and immunosuppressed group. The number of parasites was comparedstatistically using student’ t-pair test. Results showed that dexamethasone at a dose of 5.2 mg/20 g body weight intraperitoneally to the immunosuppresed mice did not increase the number of extracelluler parasites in the peritonealfluid. The best parasite harvest time was on the 4th day after inoculation. Determination of parasite protein obtainedat 4 days after inoculation using sodium dodecyl sulphate polyacrylamide gel electrophoresis showed a dominantsurface protein of 42 kDa.
PROFIL TERAPI OBAT PADA PASIEN RAWAT INAP DENGAN DIARE AKUT PADA ANAK DI RUMAH SAKIT UMUM NEGARA Arifani Siswidiasari; Ketut Widyani Astuti; Sagung Chandra Yowani
Jurnal Kimia (Journal of Chemistry) Vol. 8, No. 2 Juli 2014
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (86.374 KB) | DOI: 10.24843/JCHEM.2014.v08.i02.p07

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The Diarrhea in children is the highest cause of morbidity and mortality in the world, especially in the developing countries. The purposes of this research are to describe about patient profile, the use of medicines profile and the treatment period in the RSU Negara. The research method used was a descriptive retrospective by taking medical records of childhood diarrhea patients in The Public Hospital RSU Negara  from July to December 2012. The diarrhea type studied was acute diarrhea in children  in age range of 0-14 years. The results showed that the drugs which were used for the acute diarrhea treatment in the Public Hospital of RSU Negara were: male (69.57%), female (30.43%), with age range: 0 - <1 year old (60.87%), 1 - <5 years old (34.78%), and 5 - <14 years old (4.35%). The highest classification of acute diarrhea were acute diarrhea with mild dehydration (63.04%) and without dehydration (36.96%). The use of antibiotics (89.13%), without antibiotics (10.87%), the use of ringer's lactate (93.48%), dextrose (13.04%), zinc (65.22%), antiemetic (58.69%), antipyretic (54.35 %), antacids (2.17%), H2 blocker (23.91%), probiotics (21.74%), synbiotics (34.78%), ORS (10.87%), dexamethasone (4.35%), dygestive enzymes (2.17%), nystatin (2.17%). The condition of diarrhea patient when they were released from the hospital were cured (67.39%), begins to be cured (32.61%), with various of  treatment duration from  3 days (69.57%) , 4 days (23.91%), and 5 days (6.52%).
OPTIMASI DIGESTI ENZIM RESTRIKSI SacII PADA ISOLAT Mycobacterium tuberculosis H37Rv UNTUK DETEKSI MUTASI PROMOTER inhA PADA KASUS MDR-TB DENGAN METODE PCR-RFLP Ida Ayu Ratih Dwi Nugraha Putri; Sagung Chandra Yowani; I Nengah Wirajana
Metamorfosa: Journal of Biological Sciences Vol 4 No 1 (2017)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2017.v04.i01.p14

Abstract

The aim of this study was to optimize the digestion process of SacII restriction enzyme in Mycobacterium tuberculosis H37Rv isolate. SacII enzyme that have CCGC^GG restriction site could be used to detect the mutation at -24 promoter region of inhA M. tuberculosis. The mutation in this position is one of the causes of isoniazid resistance. The methods of this study were conducted in several step, respectively: DNA isolation of M. tuberculosis H37Rv, amplification of inhA promoter region of M. tuberculosis H37Rv using PCR method, and digest optimization using SacII. In this study, optimization of SacII digestion was conducted for formulaion and incubation time. The formulation was optimized by GM-ONE Buffer and GM-Buffer IV, meanwhile the incubation time was optimized in 1 hour, 2 hour, and 3 hour. The result of this study showed that SacII work optimally in GM-Buffer IV compared to GM-ONE Buffer and incubate for 3 hours. Under the optimal digest condition, SacII enzyme could be used to detect the mutation at -24 promoter region of inhA in the case of MDR-TB using PCR-RFLP method.
DESAIN TAQMAN PROBE SECARA IN SILICO SEBAGAI PENDETEKSI MUTASI PADA KODON 516 GEN rpoB Mycobacterium tuberculosis UNTUK METODE REAL-TIME PCR Dek Pueteri Dewi Suryani; Putu Sanna Yustiantara; Sagung Chandra Yowani
Metamorfosa: Journal of Biological Sciences Vol 4 No 1 (2017)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2017.v04.i01.p04

Abstract

The aim of this research was to in silico design of TaqMan probe. Design of TaqMan probe were conducted using software Clone Manager Suite 6. As a template, the rpoB gene of M. tuberculosis H37Rv (accession number U12205.1) was used. The results of this research were 8 sequences such as, R516MV-2, R516MV-3, R516MV-4, R516MV-5, R516MV-7, R516MV-8, R516MV-11, R516MV-13. These sequences were met the criteria of TaqMan probe, such as length of nucleotide (23-28 nucleotide), Tm value (72ºC), %GC (50-58%), runs and repeats (?4 bases), dimer structure in accordance to the requirements and does not form hairpin structures. In addition, these sequences were met labeling criteria of TaqMan probe which are including the location of G bases and the number of G-C bases in sequences. Therefore, these sequences could be labeled by FAM (reporter) at 5' end and TAMRA (quencher) at 3' end. The conclusion of this research, the sequences were met the criteria of TaqMan probe. Therefore, it could be targeted to detect mutations at codon 516 with a change of aspartic acid into valine (GAC ? GTC) by using real-time PCR method.
PEMBERDAYAAN IBU-IBU, BIDAN DAN KADER POSYANDU MELALUI PENYULUHAN TENTANG PEMBERIAN MAKANAN SEHAT UNTUK MENINGKATKAN GIZI DAN DAYA TAHAN TUBUH BAYI DAN ANAK DI DESA PENGOTAN KABUPATEN BANGLI Rini Noviyani; N. P. Ariantari; Rasmaya Niruri; K. Widyani Astuti; S. Chandra Yowani
Buletin Udayana Mengabdi Vol 13 No 2 (2014): Vol 13, No. 2 (2014)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat

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Abstract

The Community Services was held in Bangli Utara Public Health Facility, Pengotan district, Bangli. The purpose of this activity is to give information about balanced nutition and massage for babies to participants which are housewives, midwifery and posyandu staff in Bangli Utara Public Health Facility, Pengotan. They were expected to have better understanding about balanced nutrition compared to imbalanced nutrition and how to massage their babies. The methods used in this activity were deliver information about balanced nutrition and invited discussion also the baby massage practice trained to them. The result showed that participants were very interested in our program. It could be proved by their activity in discussion.Keywords: information, balanced nutrition, baby massage practice
DESAIN PRIMER SECARA IN SILICO UNTUK AMPLIFIKASI FRAGMEN GEN rpoB Mycobacterium tuberculosis DENGAN POLYMERASE CHAIN REACTION (PCR) Pradnyaniti, D.G; Wirajana, I.N; Yowani, S.C
Jurnal Farmasi Udayana Vol. 2, No. 3, Tahun 2013
Publisher : Departement of Pharmacy, Faculty of Mathematics and Natural Science, Udayana University

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Abstract

Amplifikasi DNA Mycobacterium tuberculosis dari gen rpoB dilakukan dengan metode polymerase chain reaction (PCR). Amplifikasi DNA dengan PCR diperlukan sepasang primer (forward dan reverse) untuk membatasi daerah yang ingin diamplifikasi. Penelitian ini bertujuan untuk mendesain sepasang primer agar dapat mengamplifikasi fragmen 0,5 kb gen rpoB M.tuberculosis. Desain primer dilakukan secara in silico dengan bantuan program clone manager suite 6 (University of Groningen). Template yang digunakan dalam mendesain primer adalah sekuen gen rpoB M. tuberculosis H37RV wild type, yang diperoleh dari database NCBI dengan kode genbank U12205.1. Penelitian ini telah berhasil memperoleh sekuen sepasang primer (forward dan reverse) dengan panjang masing-masing adalah 22 oligonukleotida. Primer ini dapat mengamplifikasi secara in silico fragmen 0,5 kb gen rpoB M. tuberculosis pada rentang daerah 990-1496 pb dengan panjang fragmen sebesar 507 pb.
Co-Authors A. W. Dwiputri Ade Ari Sundari Ade Ari Sundari Amelia Putri, Ni Luh Dian Saptari Arifani Siswidiasari Asmara A. A. R. Astuti, M.A.P. Ayu Nyoman Chandra Yustiana Ayu Putu Chika Iswari Anjani, Sang Cista Dewi, Ni Putu Fiona Dek Pueteri Dewi Suryani Deniariasih, N.W. Devi, Ni Wayan Antika Sri Devita Kusdianingrum Dewa Ayu Swastini Dewi Andayani Farmawati Dwicandra, N.M.O. Dyah Subadrika Warma Dewi Endang Kumolosasi Gede Bayu Surya Ekayana, I Heni Pujiastuti I Gede Ketut Sajinadiyasa I Gusti A. A. Santhi Rahmaryani I Gusti Ayu Agung Septiari I Ketut Juniarta I Ketut Tunas I Made Dira Swantara I Nengah Wirajana I Nyoman Gede Budiana I Wayan Kasa Ida Ayu Gendari Ida Ayu Ratih Dwi Nugraha Putri Ida Bagus Nyoman Putra Dwija Indra Juana Adikara Jennifer Tamara Jennifer Tamara Kadek Ratna Sari Dewi Kadek Widya Yuli Hartati Ketut Ratnayani Ketut Widyani Astuti Liangky Syane S Luh Vida Sasmitha Luk Ketut Budi Maitriani M. A. Pratiwi Made Dharmesti Wijaya Made Rai Dwitya Wiradiputra Marfu'ah, Nurul Marlia Singgih Wibowo Mirawati N.K.W. Ni Kadek Ari Cipta Pratiwi Ni Kadek Ariani Ni Ketut Sri Anggreni Ni Komang Sasi Ani Ni Made Febrianti Ni Made Yustikarini Ni Nyoman Pradnya Utari Ni Putu Ariantari Ni Putu Intan Satya Dewi Ni Putu Monica Rosdiana Dewi Paramitha Ni Wayan Sukma Pramitha Sari1 Osamu Iitsuka Pradnyaniti, D.G Putri, Ni Kadek Sri Adnyani Alit Putu Ayu Indrayathi Putu Lita Astriani Rasmaya Niruri Ratna Sari Dewi, Kadek Rini Noviyani Sang Ayu Putu Chika Iswari Anjani Saputra, Gusti Nyoman Oka Sudarma, I Wayan Ari Syamsul Arifin Tasya Pramiswari Tirtawati, Ni Putu Ayu Wijayakusuma, I Gusti Ngurah Lanang Yayanasri Yustiantara, Putu Sanna