Surface Plasmon Resonance Aptamer Biosensor for Discriminating Pathogenic Bacteria Vibrio parahaemolyticus

Abstract

In this paper, whole-bacteria SELEX (WB-SELEX) strategy was adopted to isolate specific aptamers against Vibrio parahaemolyticus. Round selection for V. parahaemolyticus was conducted 11 rounds, including two negative selection rounds. It was determined through real-time PCR amplification and post-SELEX experiment. The selected aptmers had high binding property and specificity to V. parahaemolyticus. Of 28 aptamers tested, VPCA-apta#1 had the highest binding affinity compared to other aptamer candidates obtained. To detect V. parahaemolyticus, aptamer based SPR biosensor platform was constructed and pathogenic bacteria sensing was conducted in two steps. The first step was to construct 5'-biotinylated VPCA-apta#1 binding probe. The second step was to incubate V. parahaemolyticus and test microbes in functionalized SA sensor chip in parallel. Our platform showed significant activity for detecting and discriminating V. parahaemolyticus from other enteric species such as Escherichia coli, Listeria monocytogenes, Sigella sonnei, and Vibrio fischeri. This is the first report on the use of whole-SELEX to isolate DNA aptamers specific for V. parahaemolyticus. We demonstrated the feasibility of using aptamer platform for the detection of V. parahaemolyticus in various food supplies. It might be used in multiple points of care for diagnosing Vibriosis.

Sensors, Biosensors, and Analytical Technologies for Aquaculture Water Quality

“…Table 3 is a summary of various biosensors for detecting fish pathogens. They include quartz crystal microbalance (QCM), the microcantilever sensor, amperometric sensor, potentiometric sensor, and surface plasmon resonance (SPR) biosensor, and lateral flow tests, targeting either viral RNA [92][93][94] or the bacteria cells [95][96][97][98][99]. Biosensors are typically designed to detect known bacteria or virus, but less so for identification of unknown.…”

“…To bridge the gaps of multiple parameter systems covering a wider range of parameters (physical, chemical, biochemical, and biological), firstly the chemical, biochemical, and biological sensors must reach to the desired level of portability, and then advanced WSN technology would be needed to cover not only the physical parameters but also those hard-to-measure chemical contaminants and biohazards. For all parameters, the sensor technologies must be highly robust because the aquatic environment varies from Microcantilever Vibrio cholerae O1 Cells Dynamic range 1 × 10 3 -1 × 10 7 CFU/ml LOD ∼ 1 × 10 3 CFU/ml [95] Lateral flow with AuNPs Vibrio cholerae O1 and O139 Cells LOD 10 7 cfu/ml LOD 10 3 cfu/ml after 6 h culture enrichment [96] Amperometric immunosensors Vibrio cholerae O1 Cells LOD 8 cfu/ml in seawater 80 cfu/ml in sewer water and tap water [97] Potentiometric aptasensing involving magnetic beads Vibrio alginolyticus Cells Dynamic range:10-100 cfu/ml LOD 10 cfu/ml [98] Surface plasmon resonance spectroscopy Vibrio parahaemolyticus Cells Not specified [99] 10 Research time to time. The uncontrollable environmental factor may pose challenges to the reliability of the sensing technologies.…”

Selection and applications of functional nucleic acids for infectious disease detection and prevention

“…The whole-bacteria/virus SELEX technology incorporates whole bacteria/virus with an ssDNA library by identifying oligonucleotide chains and complex markers on the bacterial surface, which does not require verification of special markers on the surface. DNA aptamers against infectious pathogens including Haemophilus influenzae type b [11], Mycobacterium tuberculosis [40], Escherichia coli (E. coli) [9,17], Streptococcus (Streptococcus pyogenes [12], Streptococcus pneumoniae [15], Staphylococcus aureus [18,41], Staphylococcus epidermidis [42]), and Vibrio (Vibrio parahaemolyticus [43], Vibrio vulnificus [10], Vibrio alginolyticus [14]) were selected. Moreover, DNA aptamers for nonpathogenic bacteria such as Bifidobacterium [13] and Vibrio fischeri [16] were selected.…”

Aptamer-based diagnostic and therapeutic approaches in animals: Current potential and challenges

“… Luo et al (2012) 49 Photoelectrochemical (PEC) immunosensor Nanoparticles based aptamer for detection of Ochratoxin A. Chen et al (2018) 50 SPR biosensor Whole bacteria detection Ahn et al (2018) 51 Biotin labeled DNA For detection of Salmonella enterica serovar Typhimurium Dwivedi et al (2013) 52 Prion protein aptamer Determination of BoIFN-γ in TB Crulhas et al (2017) 53 Specific aptamers of antibiotics Specific aptamers for detection of antibiotics such as Florfenicol, Cefquinome, Kenamycin, Oxytetracyclin, Tetracyclin and Chloramphenicol. Sadeghi et al, 2018 , Wang et al, 2018 , Xu et al, 2015 , Zhou et al, 2018 , Yan et al, 2018 …”