Soewignjo Soemohardjo
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Detection of Hepatitis B Virus Pre-core Mutant by Allele Specific Polymerase Chain Reaction Soemohardjo, Soewignjo; Widita, Haris; Muttaqin, Zainul; Gunawan, Stephanus; Wijaya, Mahendra; Wiguna, Putu Aditya; Rhamdiani, Shelly Olivia
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 14, NUMBER 1, April 2013
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Introduction: Mutation in pre-core region is characterized by negative HBeAg and positive anti-HBe despite active replications of the virus. The mutation has diagnostic and prognostic implications. Therefore, detectionof pre-core mutant is important. Standard diagnosis approach for detecting pre-core mutant is through DNA sequencing of hepatitis B virus (HBV) pre-core region. Unfortunately, DNA sequencing is not available in mostcenters. Hence, a simpler diagnostic approach is necessary.Method: An observational-analytic design study was performed. Detection of pre-core mutant was conducted in individuals with positive HBsAg and HBV DNA that had various patterns of HBeAg and anti HBe. HBsAg, HBeAg and anti-HBe was detected using immunochromatography technique. The HBV DNA was evaluated by using qualitative polymerase chain reaction (PCR) testing. PCR was done by three rounds of amplification with primers derived from wild type pre-core and mutant pre-core. Results: Of 25 sera with HBeAg negative, anti-HBe positive and HBV DNA positive, allele specific (AS) PCR pre-core mutant was detected in 20 (80%) sera. Two sera with HBeAg negative, anti HBe negative and HBV DNA positive were negative for pre-core mutant. Of 8 sera with HBeAg positive, anti HBe negative and HBV DNA positive, pre-core mutant was detected in 2 (25%) sera.Conclusion: Most of individuals with HBV DNA positive, HBeAg negative and anti-HBe positive have harbored pre-core mutant. The finding indicated that all patients with HBsAg positive, HBV DNA positive and HBeAg negative, but anti-HBe positive should be examined for the presence of pre-core mutant. Pre-core mutant is also found in HBeAg positive individual. Keywords: HBV, pre-core mutant, polymerase chain reaction
Detection of HBV-DNA and Its Correlation with the HBeAg/Anti-HBe Serological Status in HBsAg-positive Patients Widita, Haris; Soemohardjo, Soewignjo; Muttaqin, Zainul; Wiguna, Putu Aditya; Rhamdiani, Shelly Olivia; Wijaya, Mahendra; Gunawan, Stephanus
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 13, NUMBER 2, August 2012
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: In the past years, HBeAg and anti-HBe status in individuals with positive HBsAg were often correlated to viral replication. This study was aimed to find correlation between the HBV viremia and HBeAg/anti-HBe serological status in HBsAg-positive individuals. Method: An observational-analytic design was performed in this study. The sera of all positive HBsAg patients at Biomedika Hospital Laboratory were collected and examined for HBeAg and anti-HBe using immunochromatography technique between January and April 2012. The sampling method was purposive sampling. Afterwards, the sera were examined for HBV-DNA by polymerase chain reaction (PCR). Results: Sufficient amount of sera were collected from 44 patients consisting of 33 males and 11 females. The mean age was 15-68 years. Positive HBeAg and negative anti-HBe status was found in 11 (42%) patients. Negative HBeAg and positive anti-HBe was found in 26 (59.1%) patients. Both HBeAg and anti-HBe were negative in 7 (16.3%) patients. HBV-DNA was detected in all 11 (100%) patients with positive HBeAg and negative anti-HBe. HBV-DNA was also detected in 11 (42%) patients with negative HBeAg and positive anti-HBe. However, there was only one patient (14.3%) with both negative HBeAg and anti-HBe status, who had detectable HBV-DNA. Conclusion: Positive HBeAg can be used as an indicator of viremia, but negative HBeAg cannot be used as an indicator of the absence of viremia without further HBV-DNA testing. Patients with negative HBeAg and positive HBV-DNA were suspected for having pre-core mutant. Keywords: HBV-DNA, positive HBsAg, HBeAg, anti-HBe, pre-core mutant
The Detection of H pylori In Gastric Mucosal Biopsy Specimens by PCR Using Primers Derived From Ure C Gene in Patients with Dyspepsia Soemohardjo, Soewignjo; Palgunadi, I Gede; Gunawan, S; Muttaqin, Zainul; Widita, Haris; A, Wenny Astuti
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 8 ISSUE 2 August 2007
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: The detection of Helicobacter pylori (H. pylori) in gastric biopsy specimens can be done using CLO (Campylobacter Like Organism) test and histopatological examination, but the sensitivity of both Method is influenced by the density of the bacteria in the sample. Beside that, the coccoid form is detected with difficulty by histology and need immunohistochemical stain to confirm. PCR can be used for the detection of both spiral and coccoid form of the bacteria. Objective: To detect the genome of H. pylori by Polymerase Chain Reaction (PCR) using primers derived from ureC gene of the bacteria in gastric biopsy specimen from patients with dyspepsia. Methods: Gastric biopsy specimen from 179 patients with dyspepsia in the endoscopic unit Mataram hospital. The biopsy was taken from antrum and corpus and put into sterile saline for the culture of H. pylori and put into 70% ethanol solution for the PCR. The specimen for bacterial culture was carried soon to microbiology laboratory and plated into the appropriate media and grown in microaerophilic condition in CO2 incubator. The PCR was done using primers derived from ureC. Result: The H. pylori genome was detected in 79 of 179 biopsy sample (44.13%). The bacterial culture was positive for H. pylori in 22 (12%). The PCR result was positive in 10/35 of patient with normal endoscopy (28.57%). From 22 patients with duodenal ulcer without gastric ulcer the PCR was positive in 15 (68.18%). In patient with gastric ulcer without duodenal ulcer the PCR was positive in 9 patients (42.08%). From 7 patient with combined gastric and duodenal ulcer the PCR was positive in 5 (71.43%), in 3 patient with gastric cancer the PCR was positive in 1 (33.33%). Conclusion: The study showed that 44.13% of patient with dyspepsia in Mataram hospital was positive for H. pylori by PCR. Keywords: detection of Helicobacter pylori, gastric mucosal biopsy specimen, polymerase chain reaction, ureC gene
The Absence of Urease Enzymatic Activity of Helicobacter pylori Coccoid Form Jekti, Dwi Sulistya Dyah; Soemohardjo, Soewignjo; Muttaqin, Zainul
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 9, ISSUE 2, August 2008
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: Helicobacter pylori (H. pylori) is a gram negative and pleomorphic bacteria that able to change its morphology according to the environment. The objective of the study was to determine the biochemical and some genetic characteristic of coccoid form of H. pylori induced by starvation, aerobiosis and antibiotic. Method: The material of the study is an isolate of spiral form of CagA positive H. pylori grown from gastric biopsy specimen of a patient with chronic gastritis. The CagA positive isolate was subcultured in liquid media containing the sheep sera. The sample was divided into three groups each group consist of 27 tube. Each tube contained 109 CFU of H. pylori bacteria/ml in 4 ml liquid media. So the experiment was performed in 3 replicates. In the first group of sample, coccoid form was induced by a prolonged culture under microaerophilic condition without the addition of fresh media, in the second group by aerobiosis, while in the third group by addition of 0.1 µg amoxycillin/ml cultured in microaerophilic condition. Periodic sampling was done every day to calculate the percentage of coccoid form, to observe the possibility to regrow the spiral form and for serial electron microscopic observation. One tube is picked up in every periodic sampling. In tubes containing antibiotic the periodic sampling was done one hourly. Detection of cagA and ureA gene was done by Polymerase Chain Reaction (PCR) with appropriate primers. Results: The time needed for the development of coccoid form: Length of time from the start of the experiment needed to reach 100% coccoid form was: 49 days in microaerophilic with starvation, 28 days in aerobiosis with starvation, and 13.5 days in antibiotic. result of biochemical test: Urease enzymatic activity was only positive in spiral form. All samples of coccoid form due to all the 3 stressors did not show any urease enzymatic the activity. PCR of ureA gene: All samples of spiral and coccoid form showed positive band of ureA gene and cagA gene. Western blot of protein CagA, urease A and urease B: Western blot analysis showed that in spiral form and all coccoid form band of urease A and urease B is clearly seen,while cagA in Western blot only clearly seen in spiral form but it is absent in cocoid form. Conclusion: Troughout the cycle of coccoid form the urease gene responsible for the production of urease and cagA gene responsible for virulence was in intact condition. However, despite the presence of urease protein in coccoid form the urease enzymatic activity was absent. This fact has several diagnostic and clinical implications. Keywords: urease enzymatic activity, coccoid form, Helicobacter pylori
HBeAg and Anti HBe Status in Patients with Chronic Hepatitis B Infection Widita, Haris; Gunawan, Stephanus; Laksono, Baskoro Tri; Achwan, Wenny Astuti; Wilusanta, I Gusti Putu; Mahendra, Ketut; Taufiq, Herman S; Soemohardjo, Soewignjo
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 11, NUMBER 3, December 2010
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: Data on HBeAg and anti HBe status in patients with chronic hepatitis B infection are not yet available in Indonesia. This study was done to acquire data on HBeAg and anti HBe status in patients with hepatitis B chronic infection. Method: The material of this study was sera, collected from 105 patients with chronic hepatitis B infection from June to November 2007, divided into four groups of hepatoma, liver cirrhosis, chronic hepatitis B and asymptomatic HBsAg carriers. All sera were examined for HBsAg, HbeAg, anti HBe aside from liver function examinations. The sera consisted of 23 sera of patients with hepatoma, 27 with liver cirrhosis, 12 with chronic hepatitis B, and 43 with HBsAg asymptomatic carriers. Results: From 105 samples, only 18.1% samples were in replicative phase, as shown with the positivity of HBeAg and the negativity of anti-HBe. Sera with negative HbeAg and positive anti-HBe were mainly found in liver cirrhosis (70.73%) and least in chronic hepatitis B (50.00%) Conclusion: The high frequency of HBeAg negative and anti-HBe positive in this study might indicate the possible high frequency of pre core mutation. A study using quantitative HBV DNA should be done to confirm it.   Keywords: HBeAg, anti HBe, chronic hepatitis B
Correlation of CagA-Positive Strains of Helicobacter pylori with Topographic Distribution and Chronic Gastritis Grading Arinton, I Gede; Samudro, Pugud; Soemohardjo, Soewignjo; Sarjadi, Sarjadi
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 8, ISSUE 1, April 2007
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: CagA gene is a marker for the presence of Cag pathogenicity island. CagA-positive strains of Helicobacter pylori can identify individuals who have higher risk of developing gastrointestinal diseases. Aim: To discover the correlation of CagA status of Helicobacter pylori with topographic localization of Helicobacter pylori and chronic gastritis grading. Methods: Gastric biopsy specimens were taken from 104 patients. The specimens were obtained from gastric antrum, corpus and incisures for histological and polymerase chain reaction (PCR) studies. The histological chronic gastritis was assessed semi-quantitatively (grades 0-3). The PCR was used for detecting Helicobacter pylori genes and CagA strain. Topographic localization of Helicobacter pylori was classified as gastric antrum and corpus. Results: There were 33 (86.8%) CagA-positive strains of 38 patients with Helicobacter pylori-positive genes. There were no significant differences between topographic localization of Helicobacter pylori - either in the gastric antrum (rho = 0.14, p = 0.40) nor in the corpus (rho = 0.27, p =0.10) and the CagA status of Helicobacter pylori. Conclusion: CagA gene status of Helicobacter pylori does not determine chronic gastritis grading and gastric topographic localization. Keywords: chronic gastritis, cagA gene, Helicobacter pylori, gastric antrum, gastric corpus.
Detection of Hepatitis B Virus Pre-core Mutant by Allele Specific Polymerase Chain Reaction Soemohardjo, Soewignjo; Widita, Haris; Muttaqin, Zainul; Gunawan, Stephanus; Wijaya, Mahendra; Wiguna, Putu Aditya; Rhamdiani, Shelly Olivia
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 14, NUMBER 1, April 2013
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Abstract

Introduction: Mutation in pre-core region is characterized by negative HBeAg and positive anti-HBe despite active replications of the virus. The mutation has diagnostic and prognostic implications. Therefore, detectionof pre-core mutant is important. Standard diagnosis approach for detecting pre-core mutant is through DNA sequencing of hepatitis B virus (HBV) pre-core region. Unfortunately, DNA sequencing is not available in mostcenters. Hence, a simpler diagnostic approach is necessary.Method: An observational-analytic design study was performed. Detection of pre-core mutant was conducted in individuals with positive HBsAg and HBV DNA that had various patterns of HBeAg and anti HBe. HBsAg, HBeAg and anti-HBe was detected using immunochromatography technique. The HBV DNA was evaluated by using qualitative polymerase chain reaction (PCR) testing. PCR was done by three rounds of amplification with primers derived from wild type pre-core and mutant pre-core. Results: Of 25 sera with HBeAg negative, anti-HBe positive and HBV DNA positive, allele specific (AS) PCR pre-core mutant was detected in 20 (80%) sera. Two sera with HBeAg negative, anti HBe negative and HBV DNA positive were negative for pre-core mutant. Of 8 sera with HBeAg positive, anti HBe negative and HBV DNA positive, pre-core mutant was detected in 2 (25%) sera.Conclusion: Most of individuals with HBV DNA positive, HBeAg negative and anti-HBe positive have harbored pre-core mutant. The finding indicated that all patients with HBsAg positive, HBV DNA positive and HBeAg negative, but anti-HBe positive should be examined for the presence of pre-core mutant. Pre-core mutant is also found in HBeAg positive individual. Keywords: HBV, pre-core mutant, polymerase chain reaction
The Nepean Dyspepsia Index: Translation and Validation in Indonesian Language Arinton, I Gede; Samudro, Pugud; Soemohardjo, Soewignjo
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 7, ISSUE 2, August 2006
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: Dyspepsia is an important health problem from economic and quality of life point of view. However, to date, there has not been specific instrument of quality of life to evaluate patients with dyspepsia specially design in Indonesian language. The Nepean Dyspepsia Index (NDI) is a reliable and valid instrument regarding quality of life in patients with dyspepsia and had been validated in Australia, Germany, Italy and Netherlands. Objective: To report translation of NDI in Indonesian language and validation in Indonesian patients with dyspepsia and also evaluate the possibility of its use in subjects who speak Indonesian language. Methods: NDI was translated into Indonesian language. The amount of 49 subjects with a clinical diagnosis of dyspepsia according to the inclusion and exclusion criteria were recruited. Collection of data included demographic data, physical and laboratory examination. All subjects were asked to complete translation of NDI. Reliability analysis was evaluated by a-Cronbach’s and test-retest. Since dyspepsia has no gold standard, validity was evaluated using factor analysis. Result: Reliability of the questionnaire was good, a-Cronbach’s and interclass correlation coefficient were found to be > 0.70 respectively and Kaiser-Meyer-Olkin value was found to be > 0.64, suggesting that all items were appropriate to measure. Conclusion: translated NDI in Indonesian language can be used in dyspepsia, patients who understand Indonesian language. Keywords: dyspepsia, disease-related quality of life, the Nepean Dyspepsia Index, reliability, validity
The Result Discrepancies between Histological and PCR Method for Detecting Helicobacter pylori in Patients with Dyspepsia due to Inappropriate Preparation before Endoscopy Diarti, Maruni Wiwin; Widita, Haris; Soemohardjo, Soewignjo; Astuti, Weny; Sumarno, Troef; Jiwintarum, Yunan; Mutaqin, Zainul; Handayani, Retno
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 10, ISSUE 2, August 2009
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: False negative result of Helicobacter pylori (H. pylori) detection in gastric tissue can be due to inappropriate preparation before endoscopy. The objectives of this study is to compare the result of H. pylori detection in gastric biopsy by histological method and ure C polymerase chain reaction (PCR) in patients with dyspepsia who underwent upper gastrointestinal (GI) endoscopy without preparations other than six hours fasting before endoscopy. Method: We obtained 156 paraffin blocks of gastric endoscopic biopsy samples, taken from antrum and corpus of patients with dyspepsia who underwent endoscopic examination at the Endoscopy Unit of Biomedika hospital, Mataram. All biopsy samples were stained with Hematoxylin and Eosin for tissue diagnosis and Giemsa stain for detecting H. pylori Ure C PCR were done on all blocks. Cag PCR were performed on all Ure C PCR positive samples. Results: Of 156 paraffin blocks, only 17 blocks (10.9%) were positive for H. pylori by histological examination. All of the 17 samples showed positive results on PCR method. Of 156 paraffin blocks, positive results were found in 73 patients (45.9%) by ure C PCR method. The PCR method has increased the positivity rates of H. pylori more than four times compared to histological method. This study showed that the rate of cag a was 63.0%. Conclusion: Ure C PCR is superior to histological examination in patients who did not stop consuming acid supressor drug and antibiotic two weeks prior to endoscopy. This phenomenon can be explained by the change of spiral form into coccoid form of H. pylori, which is hardly detected using Giemsa stain.   Keywords: Helicobacter pylori, histology, ureC, Cag a, PCR
The Result Discrepancies between Histological and PCR Method for Detecting Helicobacter pylori in Patients with Dyspepsia due to Inappropriate Preparation before Endoscopy Diarti, Maruni Wiwin; Widita, Haris; Soemohardjo, Soewignjo; Astuti, Weny; Sumarno, Troef; Jiwintarum, Yunan; Mutaqin, Zainul; Handayani, Retno
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 10, ISSUE 2, August 2009
Publisher : The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy

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Background: False negative result of Helicobacter pylori (H. pylori) detection in gastric tissue can be due to inappropriate preparation before endoscopy. The objectives of this study is to compare the result of H. pylori detection in gastric biopsy by histological method and ure C polymerase chain reaction (PCR) in patients with dyspepsia who underwent upper gastrointestinal (GI) endoscopy without preparations other than six hours fasting before endoscopy. Method: We obtained 156 paraffin blocks of gastric endoscopic biopsy samples, taken from antrum and corpus of patients with dyspepsia who underwent endoscopic examination at the Endoscopy Unit of Biomedika hospital, Mataram. All biopsy samples were stained with Hematoxylin and Eosin for tissue diagnosis and Giemsa stain for detecting H. pylori Ure C PCR were done on all blocks. Cag PCR were performed on all Ure C PCR positive samples. Results: Of 156 paraffin blocks, only 17 blocks (10.9%) were positive for H. pylori by histological examination. All of the 17 samples showed positive results on PCR method. Of 156 paraffin blocks, positive results were found in 73 patients (45.9%) by ure C PCR method. The PCR method has increased the positivity rates of H. pylori more than four times compared to histological method. This study showed that the rate of cag a was 63.0%. Conclusion: Ure C PCR is superior to histological examination in patients who did not stop consuming acid supressor drug and antibiotic two weeks prior to endoscopy. This phenomenon can be explained by the change of spiral form into coccoid form of H. pylori, which is hardly detected using Giemsa stain.   Keywords: Helicobacter pylori, histology, ureC, Cag a, PCR