Immuno-PCR을 이용한 한국형 A형 구제역 바이러스 진단
Detection of Korean Foot-and-Mouth Disease Virus Type A by Immuno-PCR
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초록
구제역은 발생 시 살처분, 백신 접종과 같은 질병 방제 비용 발생, 생산성 감소, 동물과 축산물의 국제적인 교역 제한 등으로 국가 경제에 미치는 영향이 매우 큰 질병이다. 바이러스의 혈청형을 민감하고 정확 및 신속하게 구분할 수 있는 적합한 진단법은 구제역의 조기 예방 측면에서 지속적으로 요구되고 있다. 본 연구는 한국형 구제역 바이러스를 진단하는 방법으로, 기존 연구에서 민감성이 확인된 Immuno-PCR 진단법을 적용시킬 수 있는지를 확인하기 위해 진행되었다. Immuno-PCR을 진행하기 위해 한국에서 발병한 구제역 바이러스의 서열을 모두 확인한 뒤 적합한 서열을 선정해 항원으로 사용하기 위한 재조합 FMDV를 합성하였다. 이들을 발현시켜 항원으로 삼아 다클론 항체 및 단일클론 항체를 만들었으며, 자성입자와 올리고머를 각각 항체에 고정시켜 본 실험에 사용하였다. Western blot과 ELISA 반응과 real-time PCR 결과를 토대로 immuno-PCR을 이용하여 한국형 구제역 바이러스(A형)를 검출해낼 수 있다는 결과를 도출하였다. 연구의 결과는 본 진단법이 차후 한국형 구제역 바이러스를 더 민감하게 검출할 수 있는 진단법의 토대가 될 것이라 여겨진다.
Abstract
In the case of foot-and-mouth disease (FMD), there is a great deal of impact on the national economy due to the disposal of diseases, the cost of disease control such as vaccination, reduction of productivity, and restriction of international trade of livestock products. Therefore, appropriate diagnostic methods for sensitive, accurate and rapid identification of virus serotypes are continuously required in terms of early prevention of FMD. This study was conducted to confirm the feasibility of immuno-PCR diagnostic method for the more sensitive detection of Korean FMD virus (FMDV). We synthesized a partial FMD type A viral gene. Protein antigen, monoclonal and polyclonal antibodies of FMDV were cloned, expressed and purified and then magnetic particles were attached to polyclonal antibodies and and oligomers to monoclonal antibodies for the immnuno-PCR. We confirmed the antigen-antibody and oligomer reaction using ELISA, Western blot, and real-time PCR. These results show that Korean FMDV can be detected by using difference of Ct values between positive group and negative group using immuno-PCR.. The results of this study also suggest that this technique will be the basis of the diagnosis method to detect Korean FMDV more sensitively in the future.
참고문헌 (21)
- Brito, B. P., "Review of the global distribution of foot‐and‐mouth disease virus from 2007to 2014", Transbound. Emerg. Dis, 64(2), pp.316~332 (2015)
- Chang, L., "Immuno-PCR: An ultrasensitive immunoassay for biomolecular detection", Analytica. Chimica. Acta, 910, pp.12~24 (2016)
- Cho, H. W., "Years of epidemics (2009–2011): Pandemic influenza and foot-and-mouth disease epidemic in Korea", Osong. Public. Health. Res. Perspect, 4(3), pp.125~ (2013)
- Davies, G., "Foot and mouth disease", Res. Vet. Sci, 73(3), pp.195~199 (2002)
- Ding, Y. Z., "A highly sensitive detection for foot-and-mouth disease virus by gold nanopariticle improved immuno", Virol. J, 8(1), pp.148~ (2011)
- Grubman, M. J., "Foot-and-mouth disease", Clin. Microbiol. Rev, 17, pp.465~493 (2004)
- 곽경록, "구제역 진단법 연구개발 현황 (총설)", 동물자원연구, 28(2), pp.78~96 (2017), 동물생명과학연구소[DOI]
- Knight‐Jones, T. J. D., "Global Foot‐and‐Mouth Disease Research Update and Gap Analysis: 4‐Diagnostics", Transbound. Emerg. Dis, 63(1), pp.42~48 (2016)
- Knowles, N. J., "Southeast Asian foot-and-mouth disease viruses in Eastern Asia", Emerging. Infect. Dis, 18(3), pp.499~ (2012)
- Current Status of Foot-and-Mouth Disease and Improvement Plan, "KREI (Korea Rural Economic Institute)", Korea Rural Economic Institute, ~ (2017)
- MAFRA (Ministry of Agriculture, Food and Rural Affairs), "Foot and mouth disease standing operating procedure", ~ (2017)
- Oem, J. K., "Comparison and analysis of the complete nucleotide sequence of foot-and-mouth disease viruses from animals in Korea and other PanAsia strains", Virus. Genes, 29(1), pp.63~71 (2004)
- Park, J. H., "Control of foot-andmouth disease during 2010–2011 epidemic, South Korea", Emerging. Infect. Dis, 19(4), pp.655~ (2013)
- 박종현, "Control of type O foot-and-mouth disease by vaccination in Korea, 2014–2015", Journal of Veterinary Science, 19(2), pp.271~279 (2018), 대한수의학회[DOI]
- Paton, D. J., "Options for control of foot-and-mouth disease:knowledge, capability and policy", Philos. Trans. R. Soc. Lond. B. Biol. Sci, 364, pp.2657~2667 (2009)
- QIA (Animal and Plant Quarantine Agency), "Training Materials of Strengthen Foot-and-Mouth Disease Disinfection Management", ~ (2015)
- Rowlands, D. J., "Chemical basis of antigenic variation in foot-and-mouth disease virus", Nature, 306(5944), pp.694~ (1983)
- Sobrino, F., "Multiple genetic variants arise in the course of replication of foot-and-mouth disease virus in cell culture", Virology, 128(2), pp.310~318 (1983)
- Yang, M., "Characterization of monoclonal antibodies against foot-and-mouth disease virus serotype O and application in identification of antigenic variation in relation to vaccine strain selection", Virol. J, 11(1), pp.136~ (2014)
- Yoon, H., "Clinical manifestations of foot‐andmouth disease during the 2010", 2011 epidemic in the Republic of Korea, 2012, pp.59(6), 517~525
- Li Huachun, "Whole Genome Sequencing of A Candidate Strain for FMDV Vaccine: Genomic Structure and Genetic Variation", Molecular Pathogens (2014), Sophia Publishing Group, Inc.[DOI]