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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Low, Kim-Fatt Zain, Zainiharyati Mohd Yean, Chan Yean |
| Description | Author Affiliation: Low KF ( Faculty of Applied Sciences, Universiti Teknologi MARA, Tapah Campus, Tapah Road, Perak, 35400 Malaysia. Electronic address: lowkimfatt@perak.uitm.edu.my.); Zain ZM ( Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, Selangor, 40450 Malaysia.); Yean CY ( Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150 Malaysia. Electronic address: yeancyn@yahoo.com.) |
| Abstract | A novel enzyme/nanoparticle-based DNA biosensing platform with dual colorimetric/electrochemical approach has been developed for the sequence-specific detection of the bacterium Vibrio cholerae, the causative agent of acute diarrheal disease in cholera. This assay platform exploits the use of shelf-stable and ready-to-use (shelf-ready) reagents to greatly simplify the bioanalysis procedures, allowing the assay platform to be more amenable to point-of-care applications. To assure maximum diagnosis reliability, an internal control (IC) capable of providing instant validation of results was incorporated into the assay. The microbial target, single-stranded DNA amplified with asymmetric PCR, was quantitatively detected via electrochemical stripping analysis of gold nanoparticle-loaded latex microspheres as a signal-amplified hybridization tag, while the incorporated IC was analyzed using a simplified horseradish peroxidase enzyme-based colorimetric scheme by simple visual observation of enzymatic color development. The platform showed excellent diagnostic sensitivity and specificity (100%) when challenged with 145 clinical isolate-spiked fecal specimens. The limits of detection were 0.5ng/ml of genomic DNA and 10 colony-forming units (CFU)/ml of bacterial cells with dynamic ranges of 0-100ng/ml (R =0.992) and log (1-10 CFU/ml) (R =0.9918), respectively. An accelerated stability test revealed that the assay reagents were stable at temperatures of 4-37°C, with an estimated ambient shelf life of 200 days. The versatility of the biosensing platform makes it easily adaptable for quantitative detection of other microbial pathogens. |
| ISSN | 09565663 |
| Volume Number | 87 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2017-01-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Biosensing Techniques Methods Cholera Diagnosis Electrochemical Techniques Feces Microbiology Vibrio Cholerae Isolation & Purification Instrumentation Colorimetry Dna, Bacterial Analysis Genetics Equipment Design Gold Chemistry Humans Metal Nanoparticles Nucleic Acid Hybridization Point-of-care Systems Reproducibility Of Results Journal Article Validation Studies Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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