Ling-Shan Yu1, Sheng-Yeh Chou2, Hsing-Yu Wu3, Yu-Cheng Chen4, Yen-Hsu Chen5. 1. Division of Infectious Disease, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan. Electronic address: lingshanyu@mail.nsysu.edu.tw. 2. MobioSense Corp., Taipei, Taiwan. 3. System Manufacturing Center, National Chung-Shan Institute of Science and Technology, Taoyuan, Taiwan; Department of Electro-Optical Engineering, National Taipei University of Technology, Taipei, Taiwan. 4. Division of Infectious Disease, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 5. Department of Internal Medicine, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Institute of Graduate Medicine, Centre of Sepsis, Centre of Tropical Medicine and Infectious Diseases, Kaohsiung Medical University, Kaohsiung, Taiwan.
Abstract
BACKGROUND: African Swine Fever (ASF) is a highly contagious and lethal viral disease of swine, the presence of which in groups of pigs leads to enormous economic losses in the farming industry. However, vaccines and drugs to treat ASF have yet to be developed. To control the spread of the African Swine Fever Virus (ASFV), a diagnostic method that can be applied rapidly and can detect the disease during the early stages of infection is urgently needed. METHODS: In this study, we demonstrate a rapid and easy-to-use ASFV detection method that combines loop-mediated isothermal amplification (LAMP) and image processing with the hue-saturation-value (HSV) color model. This method was validated through use of synthetic ASFV DNA. RESULTS: The method shows high sensitivity, as it detects as few as 10 copies per reaction within 20 min. The speed and sensitivity of this newly developed assay are superior to those reported in previous studies. In addition, through HSV color space transformation, the colorimetric result of this LAMP assay can be used for a semi-quantitative analysis for ASFV (ranging from 108 to 101 copies per reaction) and improve the discern to low concentration samples from a negative control. CONCLUSION: These results show that the combination of ASFV-LAMP assay and HSV color space transformation may accelerate the screening process of pigs for ASFV infection. Overall, this study provides a rapid, sensitive, early-stage, on-site diagnosis of ASFV infection and has potential to be applied to other infectious diseases.
BACKGROUND: African Swine Fever (ASF) is a highly contagious and lethal viral disease of swine, the presence of which in groups of pigs leads to enormous economic losses in the farming industry. However, vaccines and drugs to treat ASF have yet to be developed. To control the spread of the African Swine Fever Virus (ASFV), a diagnostic method that can be applied rapidly and can detect the disease during the early stages of infection is urgently needed. METHODS: In this study, we demonstrate a rapid and easy-to-use ASFV detection method that combines loop-mediated isothermal amplification (LAMP) and image processing with the hue-saturation-value (HSV) color model. This method was validated through use of synthetic ASFV DNA. RESULTS: The method shows high sensitivity, as it detects as few as 10 copies per reaction within 20 min. The speed and sensitivity of this newly developed assay are superior to those reported in previous studies. In addition, through HSV color space transformation, the colorimetric result of this LAMP assay can be used for a semi-quantitative analysis for ASFV (ranging from 108 to 101 copies per reaction) and improve the discern to low concentration samples from a negative control. CONCLUSION: These results show that the combination of ASFV-LAMP assay and HSV color space transformation may accelerate the screening process of pigs for ASFV infection. Overall, this study provides a rapid, sensitive, early-stage, on-site diagnosis of ASFV infection and has potential to be applied to other infectious diseases.