OBJECTIVES: This paper focused on how the refractive index based nanobio-sensoring gold platform can produce more efficient, adaptable and more practical detection techniques to observe molecular interactions at high degree of sensitivity. It discusses surface chemistry approach, optimisation of the refractive index of gold platform and manipulation of gold geometry augmenting signal quality.
METHODS: In a normal-incidence reflectivity, r0 can be calculated using the Fresnel equation. Particularly at λ = 470 nm the ratio of r / r0 showed significant amplitude reduction mainly stemmed from the imaginary part of the Au refractive index. Hence, the fraction of reduction, Δr = 1 - r / r0. Experimentally, in a common reference frame reflectivity of a bare gold surface, R0 is compared with the reflectivity of gold surface in the presence of biolayer, R. The reduction rate (%) of reflectivity, ΔR = 1 - R / R0 is denoted as the AR signal. The method therefore enables quantitative measurement of the surface-bound protein by converting ΔR to the thickness, d, and subsequently the protein mass. We discussed four strategies to improve the AR signal by changing the effective refractive index of the biosensing platform. They are; a) Thickness optimisation of Au thin layer, b) Au / Ag bimetallic layer, c) composing alloy or Au composite, and d) Au thinlayer with nano or micro holes.
RESULTS: As the result we successfully 'move' the refractive index, ε of the AR platform (gold only) to ε = -0.948 + 3.455i, a higher sensitivity platform. This was done by composing Au-Ag2O composite with ratio = 1:1. The results were compared to the potential sensitivity improvement of the AR substrate using other that could be done by further tailoring the ε advanced method.
CONCLUSION: We suggested four strategies in order to realize this purpose. It is apparent that sensitivity has been improved through Au/Ag bimetallic layer or Au-Ag2O composite thin layer, This study is an important step towards fabrication of sensitive surface for detection of biomolecular interactions.
OBJECTIVES: The aim of this study is to develop a colorimetric sensor to detect Hg2+ in water sources using HRP inhibitive assay. The system can be incorporated with a mobile app to make it practical for a prompt in-situ analysis.
METHODS: HRP enzyme was pre-incubated with different concentration of Hg2+ at 37°C for 1 hour prior to the addition of chromogen. The mix of PBS buffer, 4-AAP and phenol which act as a chromogen was then added to the HRP enzyme and was incubated for 20 minutes. Alcohol was added to stop the enzymatic reaction, and the change of colour were observed and analyse using UV-Vis spectrophotometer at 520 nm wavelength. The results were then analysed using GraphPad PRISM 4 for a non-linear regression analysis, and using Mathematica (Wolfram) 10.0 software for a hierarchical cluster analysis. The samples from spectroscopy measurement were directly used for dynamic light scattering (DLS) evaluation to evaluate the changes in HRP size due to Hg2+ malfunctionation. Finally, molecular dynamic simulations comparing normal and malfunctioned HRP were carried out to investigate structural changes of the HRP using YASARA software.
RESULTS: Naked eye detection and data from UV-Vis spectroscopy showed good selectivity of Hg2+ over other metal ions as a distinctive color of Hg2+ is observed at 0.5 ppm with the IC50 of 0.290 ppm. The mechanism of Hg2+ inhibition towards HRP was further validated using a dynamic light scattering (DLS) and molecular dynamics (MD) simulation to ensure that there is a conformational change in HRP size due to the presence of Hg2+ ions. The naked eye detection can be quantitatively determined using a smartphone app namely ColorAssist, suggesting that the detection signal does not require expensive instruments to be quantified.
CONCLUSION: A naked-eye colorimetric sensor for mercury ions detection was developed. The colour change due to the presence of Hg2+ can be easily distinguished using an app via a smartphone. Thus, without resorting to any expensive instruments that are mostly laboratory bound, Hg2+ can be easily detected at IC50 value of 0.29 ppm. This is a promising alternative and practical method to detect Hg2+ in the environment.
Method: In this study, we developed a rapid, sensitive and specific insulated isothermal Polymerase Chain Reaction (iiPCR) targeting bimA gene (Burkholderia Intracellular Motility A; BPSS1492) for the identification of B. pseudomallei. A pair of novel primers: BimA(F) and BimA(R) together with a probe were designed and 121 clinical B. pseudomallei strains obtained from numerous clinical sources and 10 ATCC non-targeted strains were tested with iiPCR and qPCR in parallel.
Results: All 121 B. pseudomallei isolates were positive for qPCR while 118 isolates were positive for iiPCR, demonstrating satisfactory agreement (97.71%; 95% CI [93.45-99.53%]; k = 0.87). Sensitivity of the bimA iiPCR/POCKIT assay was 97.52% with the lower detection limit of 14 ng/µL of B. pseudomallei DNA. The developed iiPCR assay did not cross-react with 10 types of non-targeted strains, indicating good specificity.
Conclusion: This bimA iiPCR/POCKIT assay will undoubtedly complement other methodologies used in the clinical laboratory for the rapid identification of this pathogen.
METHODS: We randomly selected 500 Nepali mother-infant pairs and measured maternal intake and infant and maternal vitamin B12 status using plasma cobalamin, total plasma homocysteine, and methylmalonic acid concentrations. We revisited available children when they were 5 years old and measured growth. The associations between intake and maternal and infant markers of vitamin B12 and growth were estimated in multiple linear regression models adjusting for relevant confounders (n = 331).
RESULTS: Maternal vitamin B12 intake and status and vitamin B12 status in infancy predicted linear growth at 5 years of age, but not during infancy. Each microgram increase in the vitamin B12 intake of the mother during infancy was associated with an increase in height of 0.4 (0.2, 0.6) height-for-age z-scores and 1.7 (0.7, 2.7) cm around the child's fifth birthday.
CONCLUSION: Vitamin B12 status and intake in early life is an important determinant for linear growth at school age. Our findings should be verified in randomized, placebo controlled trials before translated into public health recommendations.