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Titis Widowati
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Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Materials Science & Nanotechnology

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Molecular Genotyping of HBV by using Nested PCR-RFLP among Hepatitis B Patients in Daerah Istimewa Yogyakarta Province and Surrounding Area Aris Haryanto; Nenny Sri Mulyani; Titis Widowati; Nastiti Wijayanti; Purnomo Hadi
Indonesian Journal of Biotechnology Vol 13, No 2 (2008)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (233.942 KB) | DOI: 10.22146/ijbiotech.7801

Abstract

Hepatitis B virus (HBV) can be classified into 8 genotypes, genotype A to H. Genotype of HBV is important for clinical and etiological investigations. Research for HBV genotyping, HBV transmission study using nested PCR and HBV genotyping based on RFLP using restriction enzymes have been reported. However, both of those methods have been not applied for HBV genotyping study among hepatitis B patients in endemic area, like Indonesia yet. Molecular genotyping of HBV will describe epidemiology, pathogenesis and clinical implication of HBV. Combination of nested PCR and RFLP (nested PCR-RFLP) method to determine HBV genotype in Indonesia is still less information. The objectives of study were to develop a system for HBV genotyping by nested PCR combined with RFLP (nested PCR-RFLP) method based on nucleotide sequence of surface protein encoding</div><div>gene (S gene) in HBV genome and to confirm HBV genotypes which predominantly found among hepatitis B patients in Daerah Istimewa Yogyakarta Province and surrounding area. Total of 149 sera from chronic hepatitis B patients from Daerah Istimewa Yogyakarta and surrounding areas were collected for in this work. Viral DNA were extracted from sera of hepatitis B patients and used as template for first round nested PCR amplification using outer primers set. Amplicons of first round PCR were used as template for second round amplification using inner primers set. Then, amplicons of second round nested PCR were restriction digested by Sty I and Bsr I enzymes. For HBV genotyping then the restriction products were analyzed by RFLP based on restriction pattern. Results showed that the first round nested PCR amplification generated DNA fragments of whole S gene in length 1.233 bp, and in second round nested PCR amplification using inner primer set generated DNA fragments 585 bp in length. Genotype analysis for all samples using nested PCR-RFLP methods by restriction digested of Sty I and Bsr I enzymes found only 2 HBV genotypes among hepatitis B patients, namely genotype B and C. Quantification</div><div>data showed that most of hepatitis B patients found infected by HBV genotype B (92,8%), genotype C (3,6%) and unidentified genotype (3,6%). Nested PCR-RFLP methods for HBV genotyping is simple and inexpensive for clinical diagnostic in large scale.
Profil dan temuan klinis pasien perdarahan saluran cerna di Departemen Kesehatan Anak RSUP Dr. Sardjito 2009 - 2015 Galih Akbar Pinandhito; Titis Widowati; Wahyu Damayanti
Sari Pediatri Vol 19, No 4 (2017)
Publisher : Badan Penerbit Ikatan Dokter Anak Indonesia (BP-IDAI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (319.986 KB) | DOI: 10.14238/sp19.4.2017.196-200

Abstract

Latar belakang. Perdarahan saluran cerna pada anak merupakan salah satu tanda bahaya sehingga diagnosis dan pengobatan dini sangat penting.Tujuan. Mengetahui etiologi dan temuan klinis perdarahan saluran cerna anak yang dirawat di RSUP Dr. Sardjito.Metode. Penelitian retrospektif menggunakan rekam medis pasien anak berusia kurang dari 18 tahun yang menjalani endoskopi dan kolonoskopi karena perdarahan saluran cerna dari Januari 2009 hingga Juni 2015. Data rekam medis yang tidak lengkap tidak diikutkan sertakan dalam penelitian.Hasil. Dari 55 pasien yang memenuhi syarat, terdapat 27 (49%) pasien perdarahan saluran cerna atas dan 28 (51%) pasien perdarahan saluran cerna bawah. Varises esofagus adalah etiologi terbanyak perdarahan saluran cerna atas 10 (37%), sedangkan untuk perdarahan saluran cerna bawah adalah kolitis 17 (60%). Perdarahan saluran pada anak lebih sering terjadi pada perempuan usia 1-6 tahun 29 (53%). Gejala yang lebih sering terjadi yaitu hematemesis 13 (24%) dan hematokezia 27 (49%). Keluhan perdarahan saluran cerna atas terbanyak adalah nyeri perut 13 (48%) sedangkan saluran cerna bawah adalah diare 8 (29%). Anemia berat ditemukan pada 5 (19%) pasien perdarahan saluran cerna atas dan 1 (4%) pasien perdarahan saluran cerna bawah.Kesimpulan. Varises esofagus merupakan penyebab tersering perdarahan saluran cerna atas sedangkan saluran cerna bawah adalah kolitis. Anak yang datang dengan keluhan nyeri perut dan diare harus dipikirkan salah satu penyebabnya adalah perdarahan saluran cerna.
Co-Authors Aris Haryanto Galih Akbar Pinandhito Nastiti Wijayanti Nenny Sri Mulyani Purnomo Hadi Wahyu Damayanti