Comparison of chemiluminescence enzyme immunoassay based on magnetic microparticles with traditional colorimetric ELISA for the detection of serum α-fetoprotein

A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).

Clinical value of chemiluminescence method for detection of antinuclear antibody profiles

BACKGROUND Antinuclear antibodies(ANAs)are crucial in diagnosing autoimmune diseases,mainly systemic lupus erythematosus(SLE).This study aimed to compare the performance of chemiluminescence assay(CLIA)and line immunoassay(LIA)in detecting ANAs in patients with autoimmune diseases,evaluate their diagnostic accuracy for SLE,and develop a novel diagnostic model using CLIA-detected antibodies for SLE.Specimens from patients with autoimmune diseases and physical examination specimens were collected to parallel detect specific antibodies.Individual antibodies'diagnostic performance and a model combining multiple antibodies were assessed.The findings provide valuable insights into improving the diagnosis of SLE through innovative approaches.AIM To compare the performance of CLIA and LIA in detecting ANAs in patients with autoimmune diseases,assess their accuracy for SLE,and develop a novel diagnostic model using CLIA-detected antibodies for SLE.METHODS Specimens have been obtained from 270 patients with clinically diagnosed autoimmune disorders,as well as 130 physical examination specimens.After that,parallel detection of anti-double-stranded DNA(dsDNA)antibody,anti-histone(Histone)antibody,anti-nucleosome(Nuc)antibody,anti-Smith(Sm)antibody,anti-ribosomal P protein(Rib-P)antibody,anti-sicca syndrome A(Ro60)antibody,anti-sicca syndrome A(Ro52)antibody,anti-sicca syndrome(SSB)antibody,anticentromere protein B(Cenp-B)antibody,anti-DNA topoisomerase 1(Scl-70)antibody,anti-histidyl tRNA synthetase(Jo-1)antibody,and anti-mitochondrial M2(AMA-M2)antibody was performed using CLIA and LIA.The detection rates,compliance rates,and diagnostic performance for SLE were compared between the two methodologies,followed by developing a novel diagnostic model for SLE.RESULTS CLIA and LIA exhibited essentially comparable detection rates for anti-dsDNA antibody,anti-Histone antibody,anti-Nuc antibody,anti-Sm antibody,anti-Rib-P antibody,anti-Ro60 antibody,anti-Ro52 antibody,anti-SSB antibody,anti-Cenp-B antibody,anti-DNAScl-70 antibody,anti-Jo-1 antibody and anti-AMA-M2 antibody(P>0.05).The two methods displayed identical results for the detection of anti-dsDNA antibody,anti-Histone antibody,anti-Nuc antibody,anti-Sm antibody,anti-Ro60 antibody,anti-Ro52 antibody,anti-SSB antibody,anti-Cenp-B antibody,anti-Scl-70 antibody,and anti-AMA-M2 antibody(Kappa>0.7,P<0.05),but showed a moderate agreement for the detection of anti-Rib-P antibody and anti-Jo-1 antibody(Kappa=0.671 and 0.665;P<0.05).In addition,the diagnostic performance of these antibodies detected by both methods was similar for SLE.The diagnostic model's area under the curve values,sensitivity,and specificity,including an anti-dsDNA antibody and an anti-Ro60 antibody detected by CLIA,were 0.997,0.962,and 0.978,respectively.These values were higher than the diagnostic performance of individual antibodies.CONCLUSION CLIA and LIA demonstrated excellent overall consistency in detecting ANA profiles.A diagnostic model based on CLIA-detected antibodies can successfully contribute to developing a novel technique for detecting SLE.

Detection and Persistence of Clinical <i>Escherichia coli</i>in Drinking Water Evaluated by a Rapid Enzyme Assay and qPCR

The aims of this study were to evaluate two methods, qPCR and a chemiluminescent assay (ColiLight II), for rapid detection of E. coli in water, and to examine the survival and persistence of clinical E. coli in drinking water and biofilm using qPCR and ColiLight II. qPCR and ColiLight II were compared with a cultivation-based method (MPN), and survival and persistence of four clinical E. coli strains in water and biofilms on stainless steel (SS) and polyethylene (PE) surfaces were studied in a flow-through reactor with non-disinfected drinking water using ColiLight II, qPCR, ATP bioluminescence, and MPN. ColiLight II and qPCR correlated well with MPN. In drinking water, some clinical E. coli strains showed prolonged survival in drinking water flow-through systems, and persisted 3 - 3.4 times longer than the theoretical washout due to incorporation into biofilms. Strain specific attributes can significantly affect detection and persistence of E. coli in drinking water matrices.

A Chemiluminescence Optical Fiber Glucose Biosensor Based on Co-immobilizing Glucose Oxidase and Horseradish Peroxidase in a Sol-gel Film

An optical fiber bienzyme sensor based on the luminol chemiluminescent reaction was developed and demonstrated to be sensitive to glucose. Glucose oxidase(GOD) and horseradish peroxidase(HRP) were co-immobilized by microencapsulation in a sol-gel film derived from tetraethyl orthosilicate(TEOS). The calibration plots for glucose were established by the optical fiber glucose sensor fabricated by attaching the bienzyme silica gel onto the glass window of the fiber bundle. The linear range was 0 2-2 mmol/L and the detection limit was approximately 0 12 mmol/L. The relative standard deviation was 5.3% ( n =6). The proposed biosensor was applied to glucose assay in ofloxacin injection successfully.

乙型肝炎表面抗原不同检测方法的优势比较分析

目的：比较化学发光微粒子免疫分析法（CMIA）和酶联免疫吸附法（ELISA）用于乙型肝炎（乙肝）表面抗原检测的优势效果。方法选取2011年3月～2013年4月采用ELISA法与CMIA法对386例患者进行了乙肝表面抗原检测的对比研究。结果 CMIA法检测乙肝表面抗原的阳性率为51.3%，ELISA法阳性率为43.5%，CMIA法阳性率明显更高，与ELISA法比较差异有统计学意义（P〈0.05）；CMIA法的灵敏度与特异性均明显高于ELISA法（P〈0.05）。结论与ELISA法比较，CMIA法对乙肝表面抗原检测在阳性率、灵敏度以及特异性等方面均存在有比较明显的优势，故建议将CMIA法作为临床检测乙肝表面抗原的首选方法而推广应用。

A Novel Enhancer for Fe^(2+)-Catalyzed Light Emission Reaction of Luminol and Dissolved Oxygen

EDTA was used as an enhancer for Fe 2+ catalyzed light emission from luminol oxidation by dissolved oxygen. As a result, the limit of detection for ferrous ion with flow injection analysis was improved by a factor of 160 by addition of EDTA to the luminol solution. Fe 2+ and Fe 3+ were determined simultaneously with a novel copper-coated zinc reductor minicolumn installed in one of the shunt after sample splitting in the manifold. The reductor minicolumn can be used for 3000 determinations at least. The dynamic range of determination was 1×10 -9 ～1×10 -5 mol·L -1 , with the limit of detection of 2.7×10 10 and 3.5×10 10 mol·L 1 ,for Fe 2+ and Fe 3+ , respectively. The preci sion for determination of 2×10 7 mol·L 1 of Fe 2+ and Fe 3+ was 2.3% and 4.0% (n=8), respectively, at a sampling rate of 60 h -1 . Cr 3+ and Co 2+ interfere. Fe 2+ and Fe 3+ in mixture were determined with satisfactory results. Samples of Fe 2+ and Fe 3+ were determined simultaneously and the results in good agreement with the standard spectrophotometric method. Indications were shown that EDTA functions as an enhancer, Fe 2+ as a catalyst, and oxygen is the oxidant of the chemiluminescent reaction, and the mechanism of the reaction was discussed.

Immunomagnetic assay combined with CdSe/ZnS amplification of chemiluminescence for the detection of abscisic acid

Abscisic acid (ABA) is an important plant hormone. It plays a key role in regulating plant responses to abiotic stress and in controlling seed germination, growth, and stomatal aperture. A rapid, sensitive analytical method for the ABA detection is urgently required for further investigation of ABA signaling. In this work, an immunomagnetic assay combined with CdSe/ZnS amplification of chemiluminescence has been developed for the detection of ABA. The result could be read out in 30 min at least, with the simplified procedure of immunomagnetic assay. Under the optimized condition, a linear range from 1 pM to 10 nM was obtained. An unexpected result induced by the dose hook effect was discussed. This method provided the high selectivity for ABA over other components that might be contained in real samples.

A NEW QUANTITATIVE DETECTION METHOD OF RECOMBINANT CFP10-ESAT6 AMALGAMATION PROTEINS FROM MYCOBACTERIUM TUBERCULOSIS BASED ON MICRO-MAGNETIC PROBES STRATEGY

A new rapid,specific and sensitive method for assay of recombinant CFP10-ESAT6 amalgamation proteins from Mycobacterium tuberculosis was proposed.The method used streptavidincoated magnetic beads to enrich the specific biotinylated anti-CFP10 antibody,then adopted a sandwich-type enzyme linked immunosorbent assay technology with two kinds of monoclonal antibodies:biotinylated anti-CFP10 antibody and HRP-labeled anti-CFP10 antibody to identify the target CFP10-ESAT6 proteins,and finally detected chemiluminescence intensity by a small home-made optical sensor.It was shown that,the corresponding chemiluminescence intensity had a good logarithmic linear response to the concentration of CFP10-ESAT6 proteins when ranging at 1~1000 ng/mL,and the correlation coefficient is 0.9937.The proposed method could detect the CFP10-ESAT6 proteins with low detection limit(1 ng/mL)and the detection time could be controlled within 45 min.Compared with commonly used detection methods of M.tuberculosis,this method was easy to operate,faster,and of higher sensitivity.The achievement of the quantitative detection of CFP10-ESAT6 proteins has important scientific significance and wide application prospects in tuberculosis control.