Ambient temperature detection of PCR amplicons with a novel sequence-specific nucleic acid lateral flow biosensor

Optimizations needed for lateral flow assay for rapid detection of pathogenic E. coli

“…In addition, the optimum amount of Ab can also be the concentration that produces a minimal decline in absorbance at λ max after the addition of salt. Results were similar to those of the studies by Ang et al (2012) using 22 µg/mL and Zhao et al (2010) using 30 µg/mL Ab to coat 40-nm GNPs. Our findings were also comparable to the study by Suria et al (2015), who defined an optimal amount as 12.0 µg/mL E. coli O157:H7 Ab for 40-nm GNPs.…”

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

“…Previously, we have described the use of a hybridisation buffer for NALF assay containing Mg 2+ cations to counteract the electrostatic repulsion between capture probes and target DNA as well as PEG to improve the hybridisation efficiency through volume exclusion (Ang et al, 2012). In the present study, amplification product generated from thermostabilised LATE-PCR mix was incompatible with the hybridisation buffer containing MgCl 2 and PEG as a decrease in signal intensity was observed when compared to that obtained with conventional LATE-PCR mix prepared without any stabilisers (Fig.…”

“…The signal generator and bio-recognition elements on the lateral flow biosensor were constructed as described previously (Ang et al, 2012). Briefly, 10 ml of colloidal gold nanoparticles (pH 8.0) were conjugated with 22 μg of anti-FITC Ab at room temperature for 1 h before passivation of gold surface was conducted with 1% (w/v) BSA for another 1 h. The pellet of colloidal gold-anti-FITC Ab conjugates was collected by centrifugation at 10,000 ×g for 30 min at 4°C and resuspended in 10 mM phosphate buffer (PB) containing 1% (w/v) BSA and 0.05% (w/v) sodium azide.…”

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

“…All LATE-PCR primers were designed according to the criteria described by Pierce et al (2005). A noncompetitive IAC primer pair flanking a 112 bp region of the thrombospondinrelated adhesive protein gene of Plasmodium falciparum (Ang et al, 2012) was incorporated into the mLATE-PCR assay to rule out false negative results.…”