Enzymatic electrochemical detection of epidemic-causing Vibrio cholerae with a disposable oligonucleotide-modified screen-printed bisensor coupled to a dry-reagent-based nucleic acid amplification assay

Vibrio cholerae and Cholera biotypes

“…Recently, Yu et al (2015) developed a nucleic acidsensing platform for detection of toxigenic Vibrio cholerae serogroups O1 and O139. The sensitivity and specificity (100%) of the platform showed excellent diagnostic capabilities when tested with 168 spiked stool samples.…”

Electrochemical-Nucleic Acid Detection with Enhanced Specificity and Sensitivity

“…This datum is collected in real time due to the magnitude and phase changing from the input to the output signal at the frequency range. To properly provide a solution for the WHO concept for new developing biosensors and answer the criteria of ASSURED [7], electrochemical DNA biosensors must be affordable. The electrochemical EIS system is a point-of-care, label free, low cost, highly sensitive method of biosensor detection [1,16].…”

“…The most common method used clinically is culture isolation of the genes to determine the microbial identification and epidemiology [3]. However, currently culture based nucleic acid detection is a lengthy process with a further lengthened period upon which treatment is administered [7]. Due to these technical limitations the World Health Organization has attempted to address this problem.…”

“…The requirements as stated by the world health organization are collectively referred to by the acronym ASSURED. ASSURED stands for the capabilities that new biosensors should have when introduced to developing countries; specifically, (i) Affordable, (ii) Specific, (iii) Sensitive, (iv) User-friendly, (v) Rapid, (vi) Equipmentfree, and (vii) deliverable to those who need it [7,8].…”

Development of a dry-reagent-based nucleic acid-sensing platform by coupling thermostabilised LATE-PCR assay to an oligonucleotide-modified lateral flow biosensor

“…The optimised dry-reagent LATE-PCR mix (20 μl) contained PCR buffer, 320 μM dNTP mix, 2.5 mM MgCl 2 , 0.5 μM ctxA_RX, 0.1 μM ctxA_FL, 0.5 μM IAC_FX, 0.05 μM IAC_RL, 5% (w/v) raffinose, 6.25% (w/v) Ficoll, 0.25 mg/ml BSA, 0.01% (w/v) orange G, 1.5 U deglycerolised Taq DNA polymerase, 10 pg of internal amplification control (IAC) template and ultrapure water. The aqueous LATE-PCR mix was lyophilised using Heto Lyolab 3000 as described by Yu et al (2015) until an amorphous glassy matrix was formed. The thermostabilised LATE-PCR mixes were stored in a desiccator or sealed in aluminium pouches containing silica gel.…”