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Molecular Surveillance of Dengue Virus Serotype Using Polymerase Chain Reaction in Surabaya 2013 Sucipto, Teguh Hari; Labiqah, Amaliah; Churrotin, Siti; Ahwanah, Nur; Mulyatno, Kris Cahyo; Soegijanto, Soegeng; Kotaki, Tomohiro; Kameoka, Masanori; Konishi, Eiji
Indonesian Journal of Tropical and Infectious Disease Vol 5, No 1 (2014)
Publisher : Institute of Topical Disease

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Dengue is one of the infectious diseases which is endemic in the tropical and sub-tropical country. The disease found in IndonesiaÂ Surabaya, 1968. The symptoms of Dengue virus infections are two kinds, first DF (Dengue Fever), second DHF (Dengue HemorrhagicÂ Fever). This infectious disease transmitted by Aedes aegypti mosquito. Mosquitoes breed in clean water areas. More than 100,000 casesÂ of DF/DHF ccurred in Indonesia every year. The purpose of this study were to provide information and the spread of dengue virusÂ types in Surabaya from January 2013 to September 2013. The nalysis technique used to determine the type of dengue virus nfectionwas used PCR (Polymerase Chain Reaction). The results obtained 69% DENV-1, 27% DENV-2 isolates, 4% isolates DENV-3, andÂ 0% DENV-4 isolates.

Differences of Universal and Multiplex Primer for Detection of Dengue Virus from Patients Suspected Dengue Hemorrhagic Fever (DHF) in Surabaya Ansori, Arif; Sucipto, Teguh; Deka, Pemta; Ahwanah, Nur; Churrotin, Siti; Kotaki, Tomohiro; Soegijanto, Soegeng
Indonesian Journal of Tropical and Infectious Disease Vol 5, No 6 (2015)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar

Dengue Hemorrhagic Fever (DHF) is a global health problem in tropical and subtropical regions, as well as endemic in110 countries around the world. Indonesia is one of the largest countries in the region of endemic dengue. In Indonesia, dengue virus infection has become a contagious disease that was very important and was reported in 1968. Many molecular epidemiological approaches have been developed to look for factor that has been assumed as the cause of the increase of prevalence dengue virus infection in the world. The aim of this study is for the detection and determination of serotype of dengue virus in Surabaya. The method used was the technique of Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Polymerase Chain Reaction (PCR) with specific primers for dengue virus. Samples suspected DHF patients were obtained from various health center and hospital in Surabaya. Results of this research detected negative result for dengue virus in all samples of patients suspected DHF. Negative results caused by dengue virus titers in serum samples of patients who had been dropped due to long storage time and taken after the third day of fever in early period.

RNA Isolation of Dengue Virus Type 2 with Different Precipitation Solvents : Methanol, Chloroform, and 2-Isopropanol. Yovilianda Maulitiva Untoro; Teguh Hari Sucipto; Harsasi Setyawati; Siti Churrotin; Ilham Harlan Amarullah; Puspa Wardhani; Aryati Aryati; Shuhai Ueda; Soegeng Soegijanto
Jurnal Kimia Riset Vol. 3 No. 1 (2018): Juni
Publisher : Universitas Airlangga

Dengue virus distributed in tropical and subtropical regions in the world. DENV viruses are transmitted between humans primarily by Aedes aegypti and Aedes albopictus mosquitoes and are endemic in most areas in which the vectors occur. Four serotypes of dengue virus are DENV-1, DENV-2, DENV-3 and DENV-4. DENV-2 is comprised of six genotypes. Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution. RNA isolation by combining Guanidinium thiocyanate and phenol reported has been reported. In this report, we investigated RNA isolation from DENV-2 using QIAamp Mini Kit with 2-Isopropanol, Methanol, Chloroform precipitation solvent. Electrophoregram showed DNA band as the result of RNA isolation with methanol and 2-isopropanol are produced quite well. Dna band of the of RNA isolation with chloroform solvent has the lowest intensity than methanol and 2-isopropanol. This study showed that methanol and 2-isopropanol can used as precipitation solvent for isolating RNA.

Precipitation Solvents for RNA Extraction of Dengue Virus Type 3: Dimethylformamide, Ethylenediamintetraacetic Acid, and Ultrapure H2O Rizqidhana Juliana Putri; Teguh Hari Sucipto; Harsasi Setyawati; Siti Churrotin; Ilham Harlan Amarullah; Puspa Wardhani; Aryati Aryati; Soegeng Soegijanto
Jurnal Kimia Riset Vol. 3 No. 2 (2018): Desember
Publisher : Universitas Airlangga

Dengue is a disease caused by a virus from the family Flaviviradae, carried by a female mosquito of Aedes aegypti species. Dengue fever is widespread in the tropic areas. It caused by rainfall, temperature and unplanned urbanization. According to the ministry of health , almost all provinces in Indonesia are endemic areas of dengue fever. In 2014, up to mid-December Dengue Hemorrhagic Fever (DHF) patients in 34 provinces in Indonesia are 71,668 people and 641. This figure is lower than the previous year, 2013 with 112,511 people and 871 deaths . This disease consists of four types of serotypes, namely DENV-1, DENV-2, DENV-3, and DENV-4. This disease can be identified using a variety of methods, one of the method is Reverse Transcription - Polymerase Chain Reaction (RT-PCR) method. This study aims to determine the ability of Dimethylformamide (DMF), Ethylenediamintetraacetic Acid (EDTA), and Ultrapure H2O as the substitute of Ethanol for precipitation in RNA extraction process. The sample used in this research obtained from Surabaya. RNA extraction itself can be done by using a special kit for RNA extraction. In Reverse Transcription - Polymerase Chain Reaction method, first RNA is extracted and then transcribed back (Reverse Transcription) which then form cDNA that later will be amplified by using PCR method. In this study used specific primers for dengue virus type 3 (DENV-3). The results of this study show that DMF, EDTA, and Ultrapure H2O can be used as the substitute of Ethanol for precipitation on RNA extraction. The result is evidenced by the formation of viral DNA bands on gel electrophoresis results.

In Vitro Study: Effect of Cobalt(II) Chloride Against Dengue Virus Type 1 in Vero Cells Teguh Hari Sucipto; Yovilianda Maulitiva Untoro; Harsasi Setyawati; Anisa Maharani; Novia Faridatus Sholihah; Siti Churrotin; Ilham Harlan Amarullah; Soegeng Soegijanto
Indonesian Journal of Pharmacy Vol 30 No 4, 2019
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjpharm30iss4pp316

Dengue virus (DENV) serotypes DENV-1 to DENV-4 are enveloped viruses that belong to the genus Flavivirus of the Flaviviridae. Dengue vaccine or antiviral has not yet been clinically approved for humans, even though there have been great efforts toward this end. Antiviral activity against DENV is needed to develop to be an alternative drug for DENV virus. Cobalt(II) chloride have been used in the treatment and prevention of diseases of humans since ancient times. The aim of this study is to investigate the antiviral effects and Cytotoxicity of Cobalt(II) chloride. This compound was further investigated for its inhibitory effect on the replication of DENV-1 in Vero cells. Antiviral activity and Cytotoxicity measured by WST-1 assay. The IC50 value of the Cobalt(II) chloride for DENV-1 was 0.38 μg/ml. The cytotoxicity of Cobalt(II) chloride to Vero cell suggest that the CC50 value was 2.91 µg/ml The results of this study demonstrate the anti-dengue serotype 1 inhibitory activity of Cobalt(II) chloride was a high toxic compound.

Molecular Surveillance of Dengue Virus Serotype Using Polymerase Chain Reaction in Surabaya 2013 Sucipto, Teguh Hari; Labiqah, Amaliah; Churrotin, Siti; Ahwanah, Nur; Mulyatno, Kris Cahyo; Soegijanto, Soegeng; Kotaki, Tomohiro; Kameoka, Masanori; Konishi, Eiji
Indonesian Journal of Tropical and Infectious Disease Vol. 5 No. 1 (2014)
Publisher : Institute of Topical Disease Universitas Airlangga

Dengue is one of the infectious diseases which is endemic in the tropical and sub-tropical country. The disease found in Indonesia Surabaya, 1968. The symptoms of Dengue virus infections are two kinds, first DF (Dengue Fever), second DHF (Dengue Hemorrhagic Fever). This infectious disease transmitted by Aedes aegypti mosquito. Mosquitoes breed in clean water areas. More than 100,000 cases of DF/DHF ccurred in Indonesia every year. The purpose of this study were to provide information and the spread of dengue virus types in Surabaya from January 2013 to September 2013. The nalysis technique used to determine the type of dengue virus nfectionwas used PCR (Polymerase Chain Reaction). The results obtained 69% DENV-1, 27% DENV-2 isolates, 4% isolates DENV-3, and 0% DENV-4 isolates.

DIFFERENCES OF UNIVERSAL AND MULTIPLEX PRIMER FOR DETECTION OF DENGUE VIRUS FROM PATIENTS SUSPECTED DENGUE HEMORRHAGIC FEVER (DHF) IN SURABAYA Ansori, Arif; Sucipto, Teguh; Deka, Pemta; Ahwanah, Nur; Churrotin, Siti; Kotaki, Tomohiro; Soegijanto, Soegeng
Indonesian Journal of Tropical and Infectious Disease Vol. 5 No. 6 (2015)
Publisher : Institute of Topical Disease Universitas Airlangga

Dengue Hemorrhagic Fever (DHF) is a global health problem in tropical and subtropical regions, as well as endemic in110 countries around the world. Indonesia is one of the largest countries in the region of endemic dengue. In Indonesia, dengue virus infection has become a contagious disease that was very important and was reported in 1968. Many molecular epidemiological approaches have been developed to look for factor that has been assumed as the cause of the increase of prevalence dengue virus infection in the world. The aim of this study is for the detection and determination of serotype of dengue virus in Surabaya. The method used was the technique of Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Polymerase Chain Reaction (PCR) with specific primers for dengue virus. Samples suspected DHF patients were obtained from various health center and hospital in Surabaya. Results of this research detected negative result for dengue virus in all samples of patients suspected DHF. Negative results caused by dengue virus titers in serum samples of patients who had been dropped due to long storage time and taken after the third day of fever in early period.

ANTIVIRAL ACTIVITY OF COPPER(II)CHLORIDE DIHYDRATE AGAINST DENGUE VIRUS TYPE-2 IN VERO CELL Sucipto, Teguh Hari; Churrotin, Siti; Setyawati, Harsasi; Kotaki, Tomohiro; Martak, Fahimah; Soegijanto, Soegeng
Indonesian Journal of Tropical and Infectious Disease Vol. 6 No. 4 (2017)
Publisher : Institute of Topical Disease Universitas Airlangga

Infection of dengue virus (DENV) was number of globally significant emerging pathogen. Antiviral dengue therapies ar importantly needed to control emerging dengue. Dengue virus (DENV) is mosquito-borne arboviruses responsible for causing acute systemic diseases and grievous health conditions in humans. To date, there is no clinically approved dengue vaccine or antiviral for humans, even though there have been great efforts towards this end. Copper and copper compounds have more effective in inactivation viruses, likes an influenza virus and human immunodeficiency virus (HIV). Purpose in this project was investigated of Copper(II)chloride Dihydrate antiviral compound were further tested for inhibitory effect on the replication of DENV-2 in cell culture. DENV replication was measures by Enzyme linked Immunosorbent Assay (ELISA) with selectivity index value (SI) was determined as the ratio of cytotoxic concentration 50 (CC50) to inhibitory concentration 50 (IC50) for compound. The maximal inhibitory concentration (IC50) of Copper(II)chloride Dihydrate against dengue virus type-2 was 0.13 Î¼g/ml. The cytotoxic concentration (CC50) of compound against Vero cell was 5.03 Î¼g/ml. The SI values for Copper(II)chloride Dihydrate 38.69. Result of this study suggest that Copper(II)chloride Dihydrate demonstated significant anti-DENV-2 inhibitory activities and not toxic in the Vero cells. Copper mechanisms play an important role in the prevention of copper toxicity, exposure to excessive levels of copper can result in a number of adverse health effects, as a result increased reactive oxygen species and oxidative damage to lipid, DNA, and proteins have been observed in human cell culture models or clinical syndromes of severe copper deficiency and inhibition was attributed to released cupric ions which react with cysteine residues on the surface of the protease.

FEVER AS INDICATOR TO SECONDARY INFECTION IN DENGUE VIRAL INFECTION Soegijanto, Soegeng; Nuryandari, Sufiandika; Churrotin, Siti; Sucipto, Teguh Hari
Indonesian Journal of Tropical and Infectious Disease Vol. 7 No. 1 (2018)
Publisher : Institute of Topical Disease Universitas Airlangga

Dengue Virus Infections are distributed in tropical and sub-tropical regions and transmitted by the mosquitoes such as Aedes aegypti and Aedes albopictus. Dengue virus can cause dengue fever, dengue hemorrhagic fever and dengue shock syndrome or dengue and severe dengue classified by World Health Organization. Beside it concurrent infection virus salmonella had been found some cases who showed fever more than 7 days. Concurrent infection with two agents can result in an illness having overlapping symptoms creating a diagnostic dilemma for treating physician, such as dengue fever with typhoid fever. The aim of this research is detection of dengue virus and secondary infection with Salmonella typhi in patients suspected dengue virus infection. Detection of dengue virus and Salmonella typhi using immunochromatography test such as NS1, IgG/IgM for dengue virus infection, and IgM/IgG Salmonella and blood culture. The fifty children with dengue virus infection came to Soerya hospital and 17 cases suspected dengue virus infection, five cases showed a positive NS1 on the second day of fever and one case concurrent with clinical manifestation of convulsi on the third days of fever there were five cases only showed positive. It was showed in this study that on the fourth to six day of fever in dengue virus infection accompanied by antibody IgM & IgG dengue. There were 12 cases showed the clinical manifestation of concurrent dengue viral infection and Salmonella, all of them showed a mild clinical manifestation and did not show plasma leakage and shock. In this study we found the length of stay of concurrent Dengue Virus Infection and Salmonella infection is more than 10 days. These patients were also more likely to have co-existing haemodynamic disturbances and bacterial septicaemia which would have required treatment with inotropes and antibiotics. This idea is very important to make update dengue viral management to decrease mortality in outbreak try to gain new prevention method before the occurrence of outbreak.

INHIBITORY ACTIVITY OF COBALT(II)–MORIN COMPLEX AGAINST THE REPLICATION OF DENGUE VIRUS TYPE 2 Sucipto, Teguh Hari; Churrotin, Siti; Setyawati, Harsasi Setyawati; Mulyatno, Kris Cahyo; Amarullah, Ilham Harlan; Ueda, Shuhai; Kotaki, Tomohiro; Sumarsih, Sri; Wardhani, Puspa; Bendryman, Sri Subekti; Aryati, Aryati; Soegijanto, Soegeng; Kameoka, Masanori
Indonesian Journal of Tropical and Infectious Disease Vol. 6 No. 6 (2017)
Publisher : Institute of Topical Disease Universitas Airlangga

Dengue virus (DENV) is a significant pathogen emerging worldwide as a cause of infectious disease. Antidengue treatments are urgently required to control the emergence of dengue. DENV is a mosquito-borne disease responsible for acute systemic diseases and serious health conditions. DENVs were distributed in the tropical and sub-tropical areas and transmitted to humans by Aedes agypty and Aedes albopictus. Dengue vaccine or antiviral has not yet been clinically approved for humans, even though there have been great efforts toward this end. Antiviral activity against DENV is an important alternative for the characterization and development of drugs. Metal–organic compounds were reported to exhibit fungicidal, bactericidal, and antiviral activities its inhibitory activity was not significant, at high concentration it was more toxic to replicating cells than to stationary cell monolayers of Vero cells. The aim of this study is to investigate the antiviral effects of Cobalt(II)–Morin complex. This compound was further investigated for its inhibitory effect on the replication of DENV-2 in Vero cells. The replication of DENV was measured by enzyme-linked immunosorbent assay and the value of selectivity index (SI). SI was determined as the ratio of the 50% cytotoxic concentration (CC50) to the 50% inhibitory concentration (IC50). The IC50 value of the Cobalt(II)–Morin complex for DENV-2 was 3.08 µg/ml, and the CC50 value of the complex for Vero cells was 3.36 µg/ml; thus, the SI value was 1.09. The results of this study demonstrate the antidengue serotype 2 inhibitory activity of Cobalt(II)–Morin complex and its high toxicity in Vero cells. Further studies are not required before Co(II)–Morin can be applied in the treatment of DENV-2 infections.

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