A novel, sensitive chemiluminescent (CL) immunoassay has been developed by taking advantage of a magnetic separation/mixing process and the amplification feature of colloidal gold label. First, the sandwich-type complex is formed in this protocol by the primary antibody immobilized on the surface of magnetic beads, the antigen in the sample, and the second antibody labeled with colloidal gold. Second, a large number of Au3+ ions from each gold particle anchored on the surface of magnetic beads are released after oxidative gold metal dissolution and then quantitatively determined by a simple and sensitive Au3+-catalyzed luminol CL reaction. Third, this protocol is evaluated for a noncompetitive immunoassay of a human immunoglobulin G, and a concentration as low as 3.1 x 10(-12) M is determined, which is competitive with colloidal gold-based anodic stripping voltammetry (ASV), colorimetric ELISA, or immunoassays based on fluorescent europium chelate labels. The high performance of this protocol is related to the sensitive CL determination of Au3+ ion (detection limit of 2 x 10(-10) M), which is 25 times higher than that by ASV at a single-use carbon-based screen-printed electrode. From the analytical chemistry point of view, this protocol will be quite promising for numerous applications in immunoassay and DNA hybridization.
The effectiveness of several zeolite catalysts was investigated using the cataluminescence (CTL) gas sensor system. Trace amounts of n-hexane in air samples were detected by this method. This research establishes that the specific pore size of the zeolite offers designable environment for selective CTL reaction, and "Lewis-type" basic sites appear to contribute to the catalytic nature of the zeolite surface. By incorporating either Cs+ or K+, the velocity and luminescence intensity of these catalytic reactions increase while going from Na to Cs, according to the basic nature of this group of cations in the following order: Cs > K > Na. The proposed sensor shows high sensitivity and selectivity to n-hexane at a mild reaction temperature of 225 degrees C. Quantitative analysis was performed at a selected wavelength of 460 nm. The linear range of CTL intensity versus concentration of n-hexane was 0.776-23.28 microg/mL (R = 0.997, n = 7) on CsNaY, and 0.776-23.28 microg/mL (R = 0.998, n = 7) on CsNaX, with a detection limit of 0.155 microg/mL (signal-to-noise ratio 3). Interferences from foreign substances such as methanol, ethanol, 2-propanol, acetone, acetonitrile, chloroform, or dichlormethane and other alkanes, aromatics, and alkyl aromatics such as methane, n-pentane, 3-methylpentane, 3,3-dimethylpentane, methylbenzene, ethylbenzene, and sec-butylbenzene were very low or not detectable. Results of a series of GC and GC/MS experiments suggest that the possible mechanism of the reaction is the formation of an unstable transition structure with a four-member ring, and this ring most probably consists of an oxygen atom and a carbonium ion localized on the zeolite suface.
Monitoring the levels of potentially toxic metal ions such as Hg(2+) in aquatic ecosystems is important because this ion can have severe effects on human health and the environment. Thus, a novel chemiluminescence (CL) sensor is developed for the highly selective and ultrasensitive detection of Hg(2+) ions in aqueous solution, based on thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination chemistry and subsequent formation of gold nanoparticles (Au NPs) in a HAuCl(4)/NH(2)OH reaction. The thus-formed Au NPs trigger the reaction between luminol and AgNO(3), producing CL emission. This novel CL technique has several advantages including high sensitivity (0.01 ppb) and selectivity over a spectrum of interfering metal ions. In view of these advantages, as well as the cost-effective, minimized working steps and portable features of the CL techniques, we expect that this CL sensor will be a promising candidate for the field detection of toxic Hg(2+) ions in environment, water, and food samples.
Here we report a new chemiluminescence reaction between basic aqueous H2O2 and acetonitrile. Its ultraweak chemiluminescence could be greatly enhanced by luminol, isoluminol-labeled streptavidin, and an Edman-type fluorescent reagent. Light emission was intense and long-lived, and this facilitated the initiation of the reaction and the measurement of the light emission. The present results permit us to propose a series of convenient, highly sensitive, and enzyme-free techniques for the detection and quantification of luminol, related conjugates, acetonitrile, and amino acids. Overall, this new chemiluminescence reaction will be quite promising for numerous applications in immunoassay, DNA hybridization, environmental monitoring. and postcolumn chromatographic detection.
There are potential advantages, in terms of simplicity and speed, for detecting DNA hybridization steps directly without using any external labels, especially for the multiplexed assays. In the current paper, we describe the use of a carrier-resolved label-free multiplexed assay for the simultaneous detection of multiple DNA targets. Herein we demonstrate that this protocol, using three homogeneous carriers thermosensitive poly(N-isopropylacrylamide), polystyrene beads, and magnetic beads, respectively, for simultaneous determination of three short DNA fragments specific to hepatitis B virus. Briefly, one hybridization occurs between a mixture of three different capture probe DNAs immobilized onto three carriers and three targets in a single vessel, and then chemiluminescence (CL) detection proceeds via an instantaneous derivatization reaction between the specific CL reagent 3,4,5-trimethoxylphenylglyoxal (TMPG) and the guanine nucleotide-rich regions within the target DNA. An excellent linearity is found within the range between 0.1 and 6.0 pmol with the lowest detection limit of 100 fmol. In contrast to current encoding strategies, every hybridization signal for the corresponding DNA target in our protocol is uniquely immobilized onto one carrier vehicle with a unique and intrinsic physical-chemical signature. Moreover, an instantaneous derivatization reaction is employed for the label-free determination of three targets in a single vessel. In addition, a simple CL setup is employed to read the carrier code instead of an expensive and complicated flow cytometer or imaging system commonly used for multiplexed assays. Further signal amplification is achieved by employing three amplified DNAs for second hybridization, which include a guanine nucleobase-rich sequence domain for the generation of light and an additional tethered nucleic acid domain complementary with one of the target DNA as an amplification platform. Such simple amplified CL transduction allows detection of DNA targets down to the 15-fmol level. This new protocol also provided a good capability in discriminating perfectly complementary DNA from single-base mismatches and noncomplementary sequences. Overall, the protocol described here may have value in a variety of clinical, environmental, and biodefense applications for which the accurate quantitative analysis of multiple DNA targets is desired.
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