Electrochemical genosensor assay using lyophilized gold nanoparticles/latex microsphere label for detection of Vibrio cholerae

Pei Sheng Liew, Benchaporn Lertanantawong, Su Yin Lee, Ravichandran Manickam, Yook Heng Lee, Werasak Surareungchai

Research output: Contribution to journalArticle

16 Citations (Scopus)

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 1 aM to 1 fM of linear target DNA. The assay was highly specific for V. cholerae, with a detection limit of 1 fM of PCR products. The ability of the sensor is to detect LAMP products as low as 50 ng μl-1. The novel lyophilized AuNPs-PSA-avidin reporter label with electrochemical genosensor detection could facilitate the rapid on-site detection of V. cholerae.

Original languageEnglish
Pages (from-to)167-173
Number of pages7
JournalTalanta
Volume139
DOIs
Publication statusPublished - 1 Jul 2015

Fingerprint

Vibrio cholerae
Latex
Microspheres
Gold
Nanoparticles
Labels
Assays
Avidin
Cholera
Sanitation
Sorbitol
DNA
Disasters
Freeze Drying
Bacteria
Gram-Negative Bacteria
Limit of Detection
Water
Sensors
Polymerase Chain Reaction

Keywords

  • Cholera
  • Genosensor
  • Gold nanoparticles
  • LAMP
  • Layer-by-layermodified latex
  • Lyophilization
  • PCR products
  • Vibrio cholerae

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Electrochemical genosensor assay using lyophilized gold nanoparticles/latex microsphere label for detection of Vibrio cholerae. / Liew, Pei Sheng; Lertanantawong, Benchaporn; Lee, Su Yin; Manickam, Ravichandran; Lee, Yook Heng; Surareungchai, Werasak.

In: Talanta, Vol. 139, 01.07.2015, p. 167-173.

Research output: Contribution to journalArticle

Liew, Pei Sheng ; Lertanantawong, Benchaporn ; Lee, Su Yin ; Manickam, Ravichandran ; Lee, Yook Heng ; Surareungchai, Werasak. / Electrochemical genosensor assay using lyophilized gold nanoparticles/latex microsphere label for detection of Vibrio cholerae. In: Talanta. 2015 ; Vol. 139. pp. 167-173.
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