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Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects.

Li P, Zhou L, Yu Y, Yang M, Ni S, Wei S, Qin Q - BMC Vet. Res. (2015)

Bottom Line: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis).Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV.Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, 510301, China.

ABSTRACT

Background: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed. 

Methods: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX).

Results: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner.

Discussion: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection.

Conclusions: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.

No MeSH data available.


Related in: MedlinePlus

QA-36 inhibited STIV infection in cultured turtles. a Cumulative mortality of turtles was recorded daily up to 10 days p.i.. QA-36 slowed the rate of STIV infection and reduced the cumulative mortality by 20 %. b Filtrates collected from ground liver and spleen tissues of dead turtles exerted a CPE in FHM cells. (Bars = 100 μm)
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Fig6: QA-36 inhibited STIV infection in cultured turtles. a Cumulative mortality of turtles was recorded daily up to 10 days p.i.. QA-36 slowed the rate of STIV infection and reduced the cumulative mortality by 20 %. b Filtrates collected from ground liver and spleen tissues of dead turtles exerted a CPE in FHM cells. (Bars = 100 μm)

Mentions: QA-36 had the highest affinity for (Fig. 3) and demonstrated the most effective inhibition of STIV (Fig. 5d), and its inhibitory effects against STIV in soft-shelled turtles were therefore studied further. Turtles injected with STIV alone showed 10 % mortality on day 3, increasing to 100 % on day 8 p.i.. In contrast, no turtles injected with the mixtures of QA-36 and STIV died until day 5 p.i. and the cumulative mortality was 80 % (Fig. 6a), suggesting that QA-36 inhibited STIV infection in vivo.Fig. 6


Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects.

Li P, Zhou L, Yu Y, Yang M, Ni S, Wei S, Qin Q - BMC Vet. Res. (2015)

QA-36 inhibited STIV infection in cultured turtles. a Cumulative mortality of turtles was recorded daily up to 10 days p.i.. QA-36 slowed the rate of STIV infection and reduced the cumulative mortality by 20 %. b Filtrates collected from ground liver and spleen tissues of dead turtles exerted a CPE in FHM cells. (Bars = 100 μm)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588899&req=5

Fig6: QA-36 inhibited STIV infection in cultured turtles. a Cumulative mortality of turtles was recorded daily up to 10 days p.i.. QA-36 slowed the rate of STIV infection and reduced the cumulative mortality by 20 %. b Filtrates collected from ground liver and spleen tissues of dead turtles exerted a CPE in FHM cells. (Bars = 100 μm)
Mentions: QA-36 had the highest affinity for (Fig. 3) and demonstrated the most effective inhibition of STIV (Fig. 5d), and its inhibitory effects against STIV in soft-shelled turtles were therefore studied further. Turtles injected with STIV alone showed 10 % mortality on day 3, increasing to 100 % on day 8 p.i.. In contrast, no turtles injected with the mixtures of QA-36 and STIV died until day 5 p.i. and the cumulative mortality was 80 % (Fig. 6a), suggesting that QA-36 inhibited STIV infection in vivo.Fig. 6

Bottom Line: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis).Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV.Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, 510301, China.

ABSTRACT

Background: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed. 

Methods: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX).

Results: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K d ) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner.

Discussion: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection.

Conclusions: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.

No MeSH data available.


Related in: MedlinePlus