Highly sensitive chemiluminescence technology for protein detection using aptamer-based rolling circle amplification platform

A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

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 significance of leukocyte chemiluminescence in patients with coronary heart diseases

In the present investigation,measurement of leukocyte chemiluminescence (CL) wasundertaken using micro-whole blood assay in 76 patients with coronary heart diseases (CHDs).The findings showed that there was a significant increase in leukocyte CL in patients with un-stable angina pectoris (UAP) and acute myocardial infarction (AM1) (P【0.01) and the in-crease was greater in AMI than that in UAP (P【0,01).It was found that leukocyte CL inAMI was correlated with the extent of the infarct and increased with the size of the infarct.Leukocyte CL in AMI patients who had received fibrinolytic treatment was markedly greaterthan that before treatment (P【0.05),suggesting that leukocytes may play an important rolein the pathogenesis of CHDs and may be involved in myocardial ischemia-reperfusion induced in-jury.The findings of the present study demonstrate that measurement of leukocyte CL may beof benefits for prognosis and treatment of CHDs.

Chemiluminescence Determination of Tetracycline and Oxytetracycline in Pharmaceutical Preparations Using Ru(bipy)_3^(2+)-Cerium(Ⅳ) System

A new chemiluminescence (CL) method for the determination of tetracycline and oxytetracycline is developed, hased on the CL reaction of tetracycline and oxytetracycline with Ru(bipy)_3~2+ and Ce(IV). In sulfuric acid medium, the CL emission is generated upon continuous oxidation of Ru(bipy)_3~2+ by cerium (IV ). The emission intensity is greatly enhanced when tetracycline and oxytetracycline are introduced into the reaction system after acid degradation. Under the optimum conditions, the calibration curves are linear over the range of 8. 0 X 10^-8^-4. 0 X 10^-6 mol/L for tetracycline and of 2. 0 X 10^-7~4. 0 X 10^-5 mol/L for oxytetracycline, with the detection limits are 4. 2 X 10^-8 mol/L for tetracycline and 1. 5 X 10^-7 mol/L for oxytetracycline, respectively. The proposed method was used for the determination of tetracycline and oxytetracycline in pharmaceutical formulations with good results.

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.

An optimized, sensitive and stable reduced graphene oxide–gold nanoparticle-luminol-H_2O_2 chemiluminescence system and its potential analytical application

The chemiluminescence （CL） performance of luminol is improved using reduced graphene oxide/gold nanoparticle （rGO-AuNP） nano-composites as catalyst. To prepare this catalyst, we propose a linker free, one-step method to in- situ synthesize rGO-AuNP nano-composites. Various measurements are utilized to characterize the resulting rGO-AuNP samples, and it is revealed that rGO could improve the stability and conductivity. Furthermore, we investigate the CL signals of luminal catalyzed by rGO-AuNP. Afterwards, the size effect of particle and the assisted enhancement effect of rGO are studied and discussed in detail. Based on the discussion, an optimal, sensitive and stable rGO-AuNP-luminon- H202 CL system is proposed. Finally, we utilize the system as a sensor to detect hydrogen peroxide and organic compounds containing amino, hydroxyl, or thiol groups. The CL system might provide a more attractive platform for various analytical devices with CL detection in the field of biosensors, bioassays, and immunosensors.

DETERMINATION OF TETRACYCLINE AND OXYTETRACYCLINE BY FLOW-INJECTION CHEMILUMINESCENCE METHOD

Objective To establish a rapid and precise continuous flow-injection chemiluminescence method for the determination of tetracycline and oxytetracycline. Methods In NaOH solution, tetracycline and oxytetracycline can sensitize obviously the chemiluminesence (CL) intensity of the reaction of luminol with KIO_4, the sensitized CL intensity is proportional to the concentration of tetracycline and oxytetracycline. So, a new flow-injection CL method has been developed. The optimum chemical conditions for the CL reaction were investigated. Results Under the optimized conditions (KIO_ 4 concentration: 1.0×10 -5 mol/L; NaOH concentration: 0.1 mol/L; luminol concentration: 1.0×10 -4mol/L), tetracycline and oxytetracycline were determined. The linear range of the working curves was 1.0×10 -7 - 1.0×10 -4g/mL, the detection limits was 1.0×10 -8g/mL and 1.1×10 -8g/mL, and the relative standard deviation was 2.6% (C_S=1.0×10 -6g/mL; n=11) and 2.0% (C_S=1.0×10 -6g/mL; n=11) respectively. Conclusion The method is simple, rapid, and sensitive, and it has been successfully applied to the the determination of tetracycline and oxytetracycline tablets, the mean recoveries being 99.7% and 98.8% respectively.

Luminol-potassium permanganate chemiluminescence system for the determination of three anthracycline antibiotics

Objective To establish a flow-injection chemiluminescence method for the determination of doxorubicin,epirubicin and mitoxantrone and study its reaction mechanism.Methods In alkaline medium,chemiluminescence of luminol-potassium permanganate system could be inhibited obviously by anthracycline antibiotics.Combined with flow-injection technique,a new chemiluminescence method for determining the anthracycline antibiotics was set up.The chemiluminescence mechanism of the luminol-potassium permanganate system was also discussed.Results Under optimal conditions,the good linear ranges of doxorubicin,epirubicin and mitoxantrone were 5.0×10-9-1.0×10-6g/mL,1.0×10-9-1.0×10-5g/mL and 3×10-9-1.0×10-6g/mL,respectively.The detection limits of doxorubicin,epirubicin and mitoxantrone were 3.0×10-9g/mL,5.0×10-8g/mL and 2.0×10-9g/mL,respectively.During eleven repeated inter-day and intra-day precision tests of 1.0×10-6g/mL samples,the relative standard deviations corresponded to reference values of 3.0%,2.8% and 2.1%.Conclusion The developed method is sensitive,accurate,rapid and of low cost.It can be applied to determine doxorubicin hydrochloride,epirubicin hydrochloride and mitoxantrone hydrochloride in injection preparations.

Determination of Biotin in Pharmaceutical Formulations by Potassium Permanganate-luminol-CdTe Nanoparticles Chemiluminescence System

A sensitive flow-injection chemiluminescence method was developed for the determination of biotin in the pharmaceutical formulations.The affinity between avidin and biotin was used to adsorb biotin on the polystyrene,with subsequent quantification of biotin based on its ability to enhance the chemiluminescence（CL） signal generated by the redox reaction of potassium permanganate-luminol-CdTe nanoparticles CL system.The investigations prove that apart from 3-aminophthalate,the CdTe quantum dots（QDs） play both catalytic and emitter roles.Under optimum conditions,the linear range for the determination of biotin was 0.01―25 ng/mL with a detection limit of 7.3×10-3 ng/mL（S/N=3）.The relative standard deviation of 5 ng/L biotin was 2.06%（n=7）.The proposed method was used to determine the biotin concentration in the pharmaceutical formulations and the recovery was between 96.4% and 104%.The proposed method is simple,convenient,rapid and sensitive.

Electrogenerated chemiluminescence sensor for the determination of metoclopramide using ordered mesoporous carbon for immobilizing tris(2,2'-bipyridyl)ruthenium

A novel electrogenerated chemiluminescence(ECL)sensor for the determination of metoclopramide was developed by employing ruthenium complex as an ECL signal producer and an ordered mesoporous carbon(OMC)material as modified material.The ECL sensor was fabricated by adsorption ruthenium complex into a mixture of OMC and Nafion,which showed good electrochemical and ECL behaviors.It was found that the ECL intensity of the sensor fabricated was greatly enhanced in the presence of metoclopramide.Based on this finding,a highly sensitive and reproducible ECL method was developed for the determination of metoclopramide.The result showed that the ECL intensity was linear with the concentration of metoclopramide in the range from 1.0×10-10 to 5.0×10-7M and the detection limit was 3×10-11M.The ECL sensor exhibited a long-term stability and a fine reproducibility with relative standard deviation of 1.0 % for 1.0×10-10M metoclopramide in 18 continuous determinations.The developed method has been applied to the determination of metoclopramide in tablet samples with satisfactory results.

Indirect Flow Injection Chemiluminescence Method for the Determination of Tetracyclines Using Cu(Ⅱ) as a Probe Ion

This paper reported an indirect flow injection chemiluminescence （FI-CL） method for the determination of the drugs tetracycline （TC）, chlortetracycline （CTC） and oxytetracycline（OTC） using Cu（ Ⅱ ） as a probe ion. The CL reaction was induced on-line and after injection of the sample the negative peaks appeared as a result of complexation. The method was applied to the determination of TCs in pharmaceuticals and human urine with recoveries in the range95-105%.

Effects of burn eschar extract on the induction of chemiluminescencein hepatic submitochondrial particles in rats

The effects of D1 , an extract of burn eschar , and D'1 , an extract of normal skin of rats , on the induction of chemiluminescense in heparocytic submitochondrial particles were studied. It was found that both D1 and D'1 were able to induce chemiluminescence in hepatocytic submitochondrial particles in a dosage-dependent manner and D1 was more potent than D'1 in this function when they were in the same dosage. When superoxide dismutase (SOD) and beta-carotene were in dosage smaller than 10μg/ml and 3. 2 mmol/L respectively, they were able to suppress the D1's capacity to induce chemiluminescence in the submitochondrial particles in a dosage-dependent manner , too. These findings suggest that there is the occurrence of free radicals in the interaction between D1' and submitochondrial particles and superoxide anions and siglet oxygen seem to be involved.

Chemiluminescence Behavior of Luminol-KIO₄-Ag Nanoparticles System and Its Analytical Applications

It was found that silver nanoparticles could strongly enhance the chemiluminescence (CL) of the luminol-KIO4 system in the presence of Co2+. The most intensive CL signals were obtained with silver nanoparticles in diameter of 22 nm. The studies of UV-visible spectra and CL spectra were carried out to explore the possible CL enhancement mechanism. Moreover, the influences of 17 amino acids and 25 organic compounds on the luminol-KIO4-Co2+-Ag NPs CL system were studied by a flow injection procedure, which led to an effective method to detect these compounds.

Evaluation of Clinical Application of Chemiluminescence and Real-time,Fluorescence-based Quantitative PCR in Diagnosis of Epstein-Barr Virus lnfection

Objective:To compare the effects of clinical application of chemiluminescence and real-time,fluorescence-based quantitative PCR in the detection Epstein-Barr virus(EBV).Methods:The data of chemiluminescence and real-time fluorescent quantitative PCR.fromipaEsfwo were suspected of being infectea w1tn rito1 roro January 2016 to January 2019 in our hospital were analyzed.The specific stage of EBV infection was analyzed,and the differences in results of the two detection methods were compared.Results:Chemiluminescence method was used to detect EBV infection during the active phase.The sensitivity of the chemiluminescence method was 76.7%(56/73)and the real-time quantitative PCRmethod was 90.4%(66/73).There was a statistical difference between the two detection methods(P<0.05).Conclusion:There was no statistical difference in positive predictive values between the chemiluminescence method and the real-time,fluorescence-based quantitative PCR method in the detection of EBV infection,but the sensitivity of chemiluminescence method is slightly lower than the real-time quantitative PCRmethod.It is noteworthy that chemiluminescence method is convenient and fast while the real-time,fluorescence-based quantitative PCR method is more accurate,which can provide a more accurate reference for clinical treatment.

Determination the Vigor of Rice Seed with Different Degrees of Aging with Ultraweak Chemiluminescence During Early Imbibition

用高灵敏度的单光子计数系统探测人工老化 (40℃ ,相对湿度 90 % ) 0、7、14和 2 1d的水稻种子吸胀初期的超弱化学发光强度变化 ,首次观测到水稻 (OryzasativaL .)种子吸胀初期超弱化学发光强度与其老化程度有关。加水激活水稻种子的超弱化学发光 ,其强度依赖于种子的老化程度。实验结果表明水稻种子的人工老化时间越短 ,则吸胀初期 (2 0～ 30min)超弱化学发光的强度越强 ,萌发率越高。水稻种子萌发率与超弱化学发光强度呈显著正相关。观测水稻种子吸胀初期超弱化学发光强度的变化可望成为检测种子老化程度的一种快速、简便、无损伤的新方法。

Chemiluminescence Determination of Benzoic Acid Using A Solid-Phase Verdigris Reactor

A new chemiluminescence flow system has been developed for sequential determina-tion of benzoic acid based on the reaction of the compound with copper carbonate entrapped in a solid-phase reactor. It was found that the unsaturated complex of Cu(II) and benzoic acid (1:1) has strong catalytic effect on the luminol-H2O2 chemiluminescence reaction. The calibration graph is linear over the range of 0.025 ~ 60 g/mL of benzoic acid, with a relative standard deviation of less than 3.0 %, and the detection limit is 0.01礸穖L-1. The proposed method was applied to the determination of benzoic acid content in different pharmaceutical formulations.

Flow injection chemiluminescence determination of meloxicam using potassium permanganate and formaldehyde system

A simple, rapid and sensitive flow injection chemiluminescence （FI-CL） method has been developed for the determination of meloxicam. The method is based on the CL-emitting reaction between meloxicam and potassium permanganate in a hydrochloric acid medium, enhanced by formaldehyde （HCHO）. Under optimum conditions, calibration curve over the range of 1.0-20.0μg/mL was obtained. The proposed method was successfully applied to the determination of meloxicam in capsules with no evi- dence of interference from common excipients. The detection limit of this method was 25.6 ng/mL. The relative standard deviation was 2.1% for 10.0 μg/mL meloxicam. The sample throughput was found to be 120 samples/h.

Study on superoxide and hydroxyl radicals generated in indirect electrochemical oxidation by chemiluminescence and UV-Visible spectra

The generation and transformation of radicals on the cathode of indirect electrochemical oxidation were studied by chemilumines- cence(CL)and UV-Visible spectra in the reactor with a salt bridge that connected the separated chambers.The CL intensity of 4×10^(-9)mol/L luminol on the cathode with bubbling oxygen was about seven times that of the intensity without it,which was because of the generation of reactive oxygen species(ROS).The existence of ROS,especially the generation of the superoxide radical,coul...

基于CX3CL1/CX3CR1信号通路探讨加味补阳还五汤对卒中后抑郁大鼠神经可塑性的影响

目的探究加味补阳还五汤对卒中后抑郁(PSD)大鼠神经可塑性的影响。方法60只大鼠随机分为假手术组、模型组、氟西汀组及加味补阳还五汤低、中、高剂量组,采用复合方法制备PSD模型,加味补阳还五汤低、中、高剂量组给予不同剂量加味补阳还五汤(6.7、13.4、26.8 g/kg)灌胃,氟西汀组给予氟西汀(1.8 mg/kg)灌胃,假手术组和模型组给予等体积蒸馏水灌胃,连续21 d。给药结束后,进行神经功能、行为学评分,HE、尼氏染色观察脑组织病理变化,免疫荧光染色观察神经元(Neun)和小胶质细胞(Iba1)之间相互作用,Western blot法检测神经趋化因子(CX3CL1)、CX3CR1、磷脂酰肌醇3激酶(PI3K)、苏氨酸激酶(Akt)蛋白表达。结果与模型组比较,各给药组神经功能缺损评分降低(P<0.05,P<0.01),旷场活动总距离和进入中央区域频次增加(P<0.05,P<0.01),不动时间缩短(P<0.05);加味补阳还五汤中、高剂量组和氟西汀组新奇摄食潜伏期缩短(P<0.01),缺血侧海马细胞排列整齐,细胞水肿减轻,细胞基本正常,尼氏小体增多,CX3CL1/Neun、CX3CR1/Iba1荧光强度增强(P<0.05),CX3CL1、CX3CR1、PI3K、p-Akt蛋白表达升高(P<0.05,P<0.01)。结论加味补阳还五汤对PSD大鼠具有抗抑郁、神经保护作用,其机制可能与调控CX3CL1/CX3CR1信号通路关键因子、增强神经可塑性有关。

Chemiluminescence detection of permanganate index (COD_(Mn)) by a luminol-KMnO_4 based reaction

A novel chemiluminescence （CL） system for determination of permanganate index （CODMn） combined with flow injection analysis has been proposed in this study. On the basis of the chemiluminescent reaction of luminol-KMnO4 system, light emission caused by luminol-KMnO4 system was detected by the photomultiplier tube, and its intensity caused by the appearance of KMnO4 after sample digestion was inversely proportional to CODMn. Effects for CODMn determining such as pH, concentrations and interference were investigated in detail. A detection limit of 0.3 mg/L CODMn with a linear range of 0.3-200 mg/L for its theoretical CODMn was obtained under the optimized experimental conditions. The relative standard deviation was 4.3% for 5.0 mg/L CODMn （n = 11）. This CL flow system for determining CODMn was simple, rapid, and suitable for automatic analysis. The data obtained by the present method were fairly in good agreement with those obtained by the standard titrimetric method. It has been applied to determine real samples with satisfactory results.