Identification and characteristics of aptamers against inactivated Vibrio alginolyticus

Zheng, Jiang; Tang, Xuemin; Wu, Renxie; Yan, Qingpi; Tang, Hua; Luo, Juan; Niu, Su‐Fang; Qu, Yue; Sun, Liwei

doi:10.1016/j.lwt.2015.07.021

Cited by 11 publications

(15 citation statements)

References 17 publications

(24 reference statements)

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“…The calculated binding affinity constants (K d ) for VA2 and VA8 were 14.31 ± 4.26 and 90.00 ± 13.51 nM, respectively (Figure ). In the previous study, other aptamers (#7, #8, #23) showed high specific binding against V. alginolyticus strain isolated in Fujian province (Zheng et al, ), then specificity of aptamers (#7, #8, #23) for V. alginolyticus strain prevalent in Beibu Gulf was also analysed. V. alginolyticus cells incubated with initial ssDNA library was served as control group (Con).…”

Section: Resultsmentioning

confidence: 99%

Selection and characterization of ssDNA aptamers specifically recognizing pathogenic Vibrio alginolyticus

Liu

et al. 2019

Journal of Fish Diseases

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Vibrio alginolyticus (V. alginolyticus) is a major opportunistic pathogen to both marine animals and humans, which has also caused heavy economic losses to mariculture. The aim of this study was to develop highly specific aptamers for V. alginolyticus. Single‐stranded DNA (ssDNA) aptamers with high binding affinity to viable V. alginolyticus were generated by Systematic Evolution of Ligands by Exponential Enrichment (SELEX) and identified by flow cytometric analysis in this study. The selected aptamers showed high specificity for V. alginolyticus and low apparent binding for other bacteria. The aptamers formed distinct stem‐loop structures, which could form the basis of aptamers’ specific binding to the target V. alginolyticus. Aptamer VA2 and VA8 showed particularly high binding affinity constant (Kd) of 14.31 ± 4.26 and 90.00 ± 13.51 nM, respectively. The aptamers produced no cytotoxic effects in vitro and in vivo. ssDNA aptamers were successfully selected against the viable bacteria pathogen V. alginolyticus by SELEX. The aptamers selected in this study could be not only applied as specific chemical molecular probes for studying V. alginolyticus pathogenesis to Trachinotus ovatus, but also developing rapid convenient diagnosis assay for V. alginolyticus infection, even when applied to the complex sample matrix, such as food and environment samples.

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Section: Resultsmentioning

confidence: 99%

Selection and characterization of ssDNA aptamers specifically recognizing pathogenic Vibrio alginolyticus

Liu

et al. 2019

Journal of Fish Diseases

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“…The aptamers against some severe aquatic viruses, such as Singapore grouper iridovirus (Li et al, ; Li, Wei, et al, ; Li, Zhou, et al, ), red‐spotted grouper nervous necrosis virus (Zhou et al, ), Trachinotus ovatus NNV (Yu, Liu, Wei, et al, ), spring viraemia of carp virus (Yu et al, ), Hirame rhabdovirus virus (Hwang et al, ), viral haemorrhagic septicaemia virus (Punnarak et al, ) and soft‐shelled turtle iridovirus (Li, Wei, et al, ; Li, Zhou, et al, ), have been successfully generated. There are also kinds of aptamers generated against various bacterial targets, such as Lactobacillus acidophilus , V. parahaemolyticus and Salmonella typhimurium (Duan, Wu, Chen, Huang, & Wang, ; Duan et al, ), V. alginolyticus (Yu, Liu, Su, et al, ; Tang, Zheng, Yan, Li, & Li, ; Zheng et al, ), V. harveyi (Zheng, Hao, Song, & Liu, ) and Aeromonas hydrophila (Li, Wang, Chen, & Li, ). Aptamers have been widely applied in different important research areas, including drug‐specific delivery (Loo et al, ; Zhao et al, ), pathogen diagnosis (Li et al, ; Li, Wei, et al, ; Li, Zhou, et al, ; Teng et al, ; Yu, Liu, Wei, et al, ; Zhou et al, ), toxin analysis (Huang et al, ; Yan et al, ) and disease pathogenesis research (Bunka & Stockley, ).…”

Section: Discussionmentioning

confidence: 99%

“…Vibrio alginolyticus ( V. alginolyticus ) is a severe opportunistic pathogen (Yu et al, ; Zhou, Hou, Li, & Qin, ). Its infection brings high mortality to maricultural animals, including sea bream, sea horse, shrimps and shellfish (Zheng et al, ). In recent years, V. alginolyticus has been frequently isolated in diseased marine cultured fish in south China and has caused heavy economic losses to mariculture (Wang, Su, & Zhang, ).…”

Section: Introductionmentioning

confidence: 99%

Development of novel aptamer‐based enzyme‐linked apta‐sorbent assay (ELASA) for rapid detection of mariculture pathogen Vibrio alginolyticus

Liu

Xiao

et al. 2019

Journal of Fish Diseases

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As the major opportunistic pathogen to both marine animals and humans, Vibrio alginolyticus (V. alginolyticus) has caused heavy economic losses to mariculture. ssDNA aptamer VA2 targeting live V. alginolyticus was generated by systematic evolution of ligands by exponential enrichment (SELEX) technology in our previous study. In this study, we first developed aptamer (VA2)‐based enzyme‐linked apta‐sorbent assay (VA2‐ELASA) for rapid detection of mariculture pathogen V. alginolyticus. The VA2‐ELASA could achieve the rapid detection for V. alginolyticus infection with high specificity and sensitivity. The VA2‐ELASA could specifically identify V. alginolyticus, but not other non‐target bacterial strains. VA2‐ELASA could detect V. alginolyticus at the concentration of 5 × 104/ml, the incubation time short to 1 min and the incubation temperature as high as 45°C, which proved sensitivity and stability of the novel VA2‐ELASA in this study. It took less than one hour to accomplish the detection process by VA2‐ELASA. The characteristics of specificity, sensitivity and easy operation make VA2‐ELASA a novel useful technology for the rapid diagnosis of pathogen V. alginolyticus in mariculture.

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“…The number of colony-forming units per mL (CFU mL −1 ) for each bacteria culture was determined by the surface plate counting method. The aptamer used in this work for the detection of V. alginolyticus was selected by Zheng et al [3]. The sequences of the aptamer DNA and other DNA are shown in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…As an opportunistic marine pathogen, Vibrio alginolyticus ( V. alginolyticus ) can not only lead to septicemias and corneal opaqueness in fish and shell diseases and white spots in shrimps [1,2,3], but also infects mammalian cells causing otitis, wound infection, and chronic diarrhea [4,5]. The rapid and sensitive detection of V. alginolyticus is of great importance for aquaculture, food industry, and clinical diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Potentiometric Aptasensing of Vibrio alginolyticus Based on DNA Nanostructure-Modified Magnetic Beads

Zhao

Ding

Han

et al. 2016

Sensors

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A potentiometric aptasensing assay that couples the DNA nanostructure-modified magnetic beads with a solid-contact polycation-sensitive membrane electrode for the detection of Vibrio alginolyticus is herein described. The DNA nanostructure-modified magnetic beads are used for amplification of the potential response and elimination of the interfering effect from a complex sample matrix. The solid-contact polycation-sensitive membrane electrode using protamine as an indicator is employed to chronopotentiometrically detect the change in the charge or DNA concentration on the magnetic beads, which is induced by the interaction between Vibrio alginolyticus and the aptamer on the DNA nanostructures. The present potentiometric aptasensing method shows a linear range of 10–100 CFU mL−1 with a detection limit of 10 CFU mL−1, and a good specificity for the detection of Vibrio alginolyticus. This proposed strategy can be used for the detection of other microorganisms by changing the aptamers in the DNA nanostructures.

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Identification and characteristics of aptamers against inactivated Vibrio alginolyticus

Cited by 11 publications

References 17 publications

Selection and characterization of ssDNA aptamers specifically recognizing pathogenic Vibrio alginolyticus

Selection and characterization of ssDNA aptamers specifically recognizing pathogenic Vibrio alginolyticus

Development of novel aptamer‐based enzyme‐linked apta‐sorbent assay (ELASA) for rapid detection of mariculture pathogen Vibrio alginolyticus

Potentiometric Aptasensing of Vibrio alginolyticus Based on DNA Nanostructure-Modified Magnetic Beads

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