Kuitio Chakpetch, Rasri Natchaya, Kiriwan Duangnapa, Unajak Sasimanas, Choowongkomon Kiattawee
Genetic Engineering Interdisciplinary Program, Graduate School, Kasetsart University, Bangkok 10900, Thailand.
Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
J Vet Sci. 2020 Sep;21(5):e79. doi: 10.4142/jvs.2020.21.e79.
Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential.
To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig.
New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM).
Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 10 particles.
SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.
最近，泰国的猪肉产业面临高致病性猪繁殖与呼吸综合征病毒（PRRSV）疫情，该病毒在包括老挝人民民主共和国和柬埔寨在内的整个东南亚地区传播。因此，在农场对感染猪进行快速现场筛查至关重要。
开发新型适配体作为检测PRRSV的生物传感器，用于对感染猪进行快速现场筛查。
采用毛细管电泳、金纳米颗粒比色法和石英晶体微天平（QCM）等技术相结合的方法，筛选出针对PRSSV的新型适配体。
通过毛细管电泳指数富集配体系统进化技术（SELEX），从PRRSV中鉴定出36个候选适配体。比色法显示，36个适配体中只有8个能与PRSSV结合。在测试的8个适配体中，只有1F适配体在存在经典猪瘟病毒和伪狂犬病病毒时能特异性结合PRSSV。利用QCM确认了1F适配体的特异性和灵敏度，检测限为 $1.87×10$ 个颗粒。
配备毛细管电泳的适配体SELEX筛选可能揭示出检测PRRSV的有前景的候选物。1F适配体对PRRSV表现出最高的特异性和选择性。这些发现表明，对于进一步开发新型PRRSV快速检测试剂盒而言，1F是一种有前景的适配体。
