Journal of veterinary science · 2020
Development of a biosensor from aptamers for detection of the porcine reproductive and respiratory syndrome virus.
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Kuitio Chakpetch, Rasri Natchaya, Kiriwan Duangnapa, Unajak Sasimanas, Choowongkomon Kiattawee
Аннотация
BACKGROUND: 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.
OBJECTIVES: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig.
METHODS: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM).
RESULTS: 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 × 1010 particles.
CONCLUSIONS: 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.
OBJECTIVES: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig.
METHODS: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM).
RESULTS: 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 × 1010 particles.
CONCLUSIONS: 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.