Affiliations 

  • 1 Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Semeling, Kedah, Malaysia
  • 2 Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkhuntien-chaitalay Road, Thakam, Bangkok 10150, Thailand. Electronic address: l_benjap@yahoo.com
  • 3 School of Chemical Sciences and Food Technology, Faculty of Science and Technology, University Kebangsaan Malaysia, Bangi, Selangor 43600, Malaysia
  • 4 Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkhuntien-chaitalay Road, Thakam, Bangkok 10150, Thailand; School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkhuntien-chaitalay Road, Thakam, Bangkok 10150, Thailand
Talanta, 2015 Jul 1;139:167-73.
PMID: 25882423 DOI: 10.1016/j.talanta.2015.02.054

Abstract

Vibrio cholerae is a Gram-negative bacterium that causes cholera, a diarrheal disease. Cholera is widespread in poor, under-developed or disaster-hit countries that have poor water sanitation. Hence, a rapid detection method for V. cholerae in the field under these resource-limited settings is required. In this paper, we describe the development of an electrochemical genosensor assay using lyophilized gold nanoparticles/latex microsphere (AuNPs-PSA) reporter label. The reporter label mixture was prepared by lyophilization of AuNPs-PSA-avidin conjugate with different types of stabilizers. The best stabilizer was 5% sorbitol, which was able to preserve the dried conjugate for up to 30 days. Three methods of DNA hybridization were compared and the one-step sandwich hybridization method was chosen as it was fastest and highly specific. The performance of the assay using the lyophilized reagents was comparable to the wet form for detection of 1aM to 1fM of linear target DNA. The assay was highly specific for V. cholerae, with a detection limit of 1fM of PCR products. The ability of the sensor is to detect LAMP products as low as 50ngµl(-1). The novel lyophilized AuNPs-PSA-avidin reporter label with electrochemical genosensor detection could facilitate the rapid on-site detection of V. cholerae.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.