• Title/Summary/Keyword: Dengue

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Development of Nucleic Acid Lateral Flow Immunoassay for Rapid and Accurate Detection of Chikungunya Virus in Indonesia

  • Ajie, Mandala;Pascapurnama, Dyshelly Nurkartika;Prodjosoewojo, Susantina;Kusumawardani, Shinta;Djauhari, Hofiya;Handali, Sukwan;Alisjahbana, Bachti;Chaidir, Lidya
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1716-1721
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    • 2021
  • Chikungunya fever is an arboviral disease caused by the Chikungunya virus (CHIKV). The disease has similar clinical manifestations with other acute febrile illnesses which complicates differential diagnosis in low-resource settings. We aimed to develop a rapid test for CHIKV detection based on the nucleic acid lateral flow immunoassay technology. The system consists of a primer set that recognizes the E1 region of the CHIKV genome and test strips in an enclosed cassette which are used to detect amplicons labeled with FITC/biotin. Amplification of the viral genome was done using open-source PCR, a low-cost open-source thermal cycler. Assay performance was evaluated using a panel of RNA isolated from patients' blood with confirmed CHIKV (n = 8) and dengue virus (n = 20) infection. The open-source PCR-NALFIA platform had a limit of detection of 10 RNA copies/ml. The assay had a sensitivity and specificity of 100% (95% CI: 67.56% - 100%) and 100% (95% CI: 83.89% - 100%), respectively, compared to reference standards of any positive virus culture on C6/36 cell lines and/or qRT-PCR. Further evaluation of its performance using a larger sample size may provide important data to extend its usefulness, especially its utilization in the peripheral healthcare facilities with scarce resources and outbreak situations.

Effect of Copper on Lethal Dose and Pupation of Aedes albopictus (Diptera: Culicidae) (흰줄숲모기, Aedes albopictus (Diptera: Culicidae)의 치사농도 및 용화에 대한 구리의 영향)

  • Lee, Su-Mi;Yoon, Su-Jin;Shin, Byung-Sik
    • Korean Journal of Environmental Biology
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    • v.28 no.2
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    • pp.64-68
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    • 2010
  • The mosquito, Aedes albopictus is important vector of humen phatogen such as dengue fever. And Aedes albopictus has widly distribution in oriental regions. This study was conducted to investigate the biological effects of cooper on the development of the Asian tiger mosquitoes, Aedes albopictus (Skuse). Aedes albopictus was affected with pupation ratios, mortalities, and wing lengths of the 3th and 4th instars by exposure to different concentration $CuCl_2$ (0.0, 2.5, 25.0 and 50.0 ppm) for 24 h and 48 h depending on dose and exposure period. The lowest $LC_{50}$ (median lethal concentration) value of $CuCl_2$ was 18.1 ppm at the 3rd instars for 48 h exposure. The pupation ratio of the larvae exposed at 50.0 ppm for 48 h was 14% which was much lower than those exposed to other treatment groups Changes of adult wing length showed significant decreased by Cu treatment. In conclusion, the most prominent effects of high concentration and longer exposure period were to reduce in the survival and pupation rates of larvae and wing length of Ae. albopictus.

Discrimination of Flaviviruses with High Frequency of Infection in Asian Countries: Epitope Prediction by Bioinformatic Approaches (아시아 국가 내에서 감염빈도가 높은 플라비바이러스의 구별: 생물정보학적 접근을 통한 항원결정기 예측)

  • Choi, Jae-Won;Jo, Byung-Gwan;Kim, Min Jung;Park, Suji;Kim, Hak Yong
    • The Journal of the Korea Contents Association
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    • v.18 no.4
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    • pp.99-113
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    • 2018
  • Recently, global warming has widened the habitat of mosquitoes and infection chances for mosquito-borne diseases are increasing. Flavivirus is a typical mosquito-borne virus. Flaviviruses with a relatively high frequency of infection in Asian countries include Zika, Dengue, and Japanese encephalitis viruses. Although distinctive diagnosis of flaviviruses is required because the symptoms and therapeutic method differ, there is no diagnostic method that can distinguish them accurately yet. In this study, we propose distinctive diagnosis method of flaviviruses using informations and analysis tools constructed in bioinformatic databases. The envelope protein and non-structural protein 1 which are useful protein for the immuno-diagnostics of three flaviviruses were selected. Their homology was analyzed by multiple sequence alignments and epitope candidates consisting of 10-15 amino acids were selected. Finally two epitopes were suggested to be most useful by immunogenicity analysis and 3D structure prediction. These approaches and results are expected to be great value in the distinctive diagnosis of three flaviviruses with a high frequency of infection in Asian countries.

Simple, Rapid and Sensitive Portable Molecular Diagnosis of SFTS Virus Using Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP)

  • Baek, Yun Hee;Cheon, Hyo-Soon;Park, Su-Jin;Lloren, Khristine Kaith S.;Ahn, Su Jeong;Jeong, Ju Hwan;Choi, Won-Suk;Yu, Min-Ah;Kwon, Hyeok-il;Kwon, Jin-Jung;Kim, Eun-Ha;Kim, Young-il;Antigua, Khristine Joy C.;Kim, Seok-Yong;Jeong, Hye Won;Choi, Young Ki;Song, Min-Suk
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1928-1936
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    • 2018
  • Recently, human infections caused by severe fever with thrombocytopenia syndrome virus (SFTSV), which can lead to fatality, have dramatically increased in East Asia. With the unavailability of vaccines or antiviral drugs to prevent and/or treat SFTSV infection, early rapid diagnosis is critical for prevention and control of the disease. Here, we report the development of a simple, rapid and sensitive portable detection method for SFTSV infection applying reverse transcription-loop mediated isothermal amplification (RT-LAMP) combined with one-pot colorimetric visualization and electro-free reaction platform. This method utilizes a pocket warmer to facilitate diagnosis in a resource-limited setting. Specific primers were designed to target the highly-conserved region of L gene of SFTSV. The detection limit of the RT-LAMP assay was approximately $10^0$ viral genome copies from three different SFTSV strains. This assay exhibited comparable sensitivity to qRT-PCR and 10-fold more sensitivity than conventional RT-PCR, with a rapid detection time of 30 to 60 minutes. The RT-LAMP assay using SFTSV clinical specimens has demonstrated a similar detection rate to qRT-PCR and a higher detection rate compared to conventional RT-PCR. Moreover, there was no observed cross-reactive amplification of other human infectious viruses including Japanese Encephalitis Virus (JEV), Dengue, Enterovirus, Zika, Influenza and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). This highly sensitive, electro- and equipment-free rapid colorimetric visualization method is feasible for resource-limited SFTSV field diagnosis.

Association of Estrogen Receptor Alpha and Interleukin 6 Polymorphisms with Lymphovascular Invasion, Extranodal Extension, and Lower Disease-Free Survival in Thai Breast Cancer Patients

  • Sa-Nguanraksa, Doonyapat;Suntiparpluacha, Monthira;Kulprom, Anchalee;Kummalue, Tanawan;Chuangsuwanich, Tuenjai;Avirutnan, Panissadee;O-Charoenrat, Pornchai
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2935-2940
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    • 2016
  • Breast cancer is the most frequent type of cancer diagnosed among women worldwide and also in Thailand. Estrogen and estrogen receptors exert important roles in its genesis and progression. Several cytokines have been reported to be involved in the microenvironment that promotes distant metastasis via modulation of immune and inflammatory responses to tumor cells. Estrogen receptor genetic polymorphisms and several cytokines have been reported to be associated with breast cancer susceptibility and aggressiveness. To investigate roles of genetic polymorphisms in estrogen receptor alpha (ESR1) and interleukin 6 (IL6), breast cancer patients and control subjects were recruited from the Division of Head, Neck and Breast Surgery (Siriraj Hospital, Bangkok, Thailand). Polymorphisms in ESR1 (rs3798577) and IL6 (rs1800795 and rs1800797) were evaluated by real-time PCR in 391 breast cancer patients and 79 healthy controls. Associations between genetic polymorphisms and clinicopathological data were determined. There was no association between genetic polymorphisms and breast cancer susceptibility. However the ESR1 rs3798577 CT genotype was associated with presence of lymphovascular invasion (OR=2.07, 95%CI 1.20-3.56, p=0.009) when compared to the TT genotype. IL6 rs1800795 CC genotype was associated with presence of extranodal extension (OR= 2.30, 95%CI 1.23-4.31, p=0.009) when compared to the GG genotype. Survival analysis showed that IL6 rs1800797 AG or AA genotypes were associated with lower disease-free survival. These findings indicate that polymorphisms in ESR1 and IL6 contribute to aggressiveness of breast cancer and may be used to identify high risk patients.