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Selective Amplification of Plasmonic Sensor Signal for Cortisol Detection Using Gold Nanoparticles.

ABSTRACT: Herein, gold nanoparticles (AuNP)-modified cortisol-imprinted (AuNP-MIP) plasmonic sensor was developed for signal amplification and real-time cortisol determination in both aqueous and complex solutions. Firstly, the sensor surfaces were modified with 3-(trimethoxylyl)propyl methacrylate and then pre-complex was prepared using the functional monomer N-methacryloyl-L-histidine methyl ester. The monomer solution was made ready for polymerization by adding 2-hydroxyethyl methacrylate to ethylene glycol dimethacrylate. In order to confirm the signal enhancing effect of AuNP, only cortisol-imprinted (MIP) plasmonic sensor was prepared without AuNP. To determine the selectivity efficiency of the imprinting process, the non-imprinted (AuNP-NIP) plasmonic sensor was also prepared without cortisol. The characterization studies of the sensors were performed with atomic force microscopy and contact angle measurements. The kinetic analysis of the AuNP-MIP plasmonic sensor exhibited a high correlation coefficient (R² = 0.97) for a wide range (0.01-100 ppb) with a low detection limit (0.0087 ppb) for cortisol detection. Moreover, the high imprinting efficiency (k' = 9.67) of the AuNP-MIP plasmonic sensor was determined by comparison with the AuNP-NIP plasmonic sensor. All kinetic results were validated and confirmed by HPLC.

SUBMITTER: Yılmaz GE

PROVIDER: S-EPMC9313393 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Acute hepatopancreatic necrosis disease (AHPND) is a component cause of early mortality syndrome (EMS) of shrimp. In 2013, the causative agent was found to be unique isolates of Vibrio parahaemolyticus (VPAHPND) that contained a 69 kbp plasmid (pAP1) carrying binary Pir-like toxin genes PirvpA and PirvpB. In Thailand, AHPND was first recognized in 2012, prior to knowledge of the causative agent, and it subsequently led to a precipitous drop in shrimp production. After VPAHPND was characterized, a major focus of the AHPND control strategy was to monitor broodstock shrimp and post larvae for freedom from VPAHPND by nucleic acid amplification methods, most of which required use of expensive and sophisticated equipment not readily available in a shrimp farm setting. Here, we describe a simpler but equally sensitive approach for detection of VPAHPND based on loop-mediated isothermal amplification (LAMP) combined with unaided visual reading of positive amplification products using a DNA-functionalized, ssDNA-labled nanogold probe (AuNP). The target for the special set of six LAMP primers used was the VPAHPND PirvpA gene. The LAMP reaction was carried out at 65°C for 45 min followed by addition of the red AuNP solution and further incubation at 65°C for 5 min, allowing any PirvpA gene amplicons present to hybridize with the probe. Hybridization protected the AuNP against aggregation, so that the solution color remained red upon subsequent salt addition (positive test result) while unprotected AuNP aggregated and underwent a color change from red to blue and eventually precipitated (negative result). The total assay time was approximately 50 min. The detection limit (100 CFU) was comparable to that of other commonly-used methods for nested PCR detection of VPAHPND and 100-times more sensitive than 1-step PCR detection methods (104 CFU) that used amplicon detection by electrophoresis or spectrophotometry. There was no cross reaction with DNA templates derived from non-AHPND bacteria commonly found in shrimp ponds (including other Vibrio species). The new method significantly reduced the time, difficulty and cost for molecular detection of VPAHPND in shrimp hatchery and farm settings.

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Selective Amplification of Plasmonic Sensor Signal for Cortisol Detection Using Gold Nanoparticles.

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