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Washing-free chemiluminescence immunoassay for rapid detection of cardiac troponin Ⅰ in whole blood samples

Chinese Chemical Letters 2022年第2期33卷 743-746页

作者： Huan Zhao Enben Su Li Huang Yunfeng Zai Yuan Liu Zhu Chen Song Li Lian Jin Yan Deng Nongyue He State Key Laboratory of Bioelectronics National Demonstration Center for Experimental Biomedical Engineering EducationSchool of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Getein Biotechnology Co. Ltd.Nanjing 210000China Hunan Key Laboratory of Biomedical Nanomaterials and Devices Hunan University of TechnologyZhuzhou 412007China

chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.

关键词： Washing-free Whole blood chemiluminescence immunoassay Polychloromethylstyrene microspheres

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Portable System for Rapid Field Test of CEA in Human Serum Based on chemiluminescence Enzyme immunoassay

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Chinese Journal of Electronics 2017年第1期26卷 152-155页

作者： LIU Juntao QU Shuxue LUO Jinping WANG Bin CAI Xinxia State Key Laboratory of Transducer Technology Institute of Electronics Chinese Academy of Sciences University of Chinese Academy of Sciences

Carcinoembryonic antigen(CEA) is one of the most widely used tumor markers worldwide. A portable system was designed for rapid field specific test of CEA based on immunomagnetic separation and chemiluminescence immunoassay, which consisted of chemiluminescent immunosensor and optical test instrument. The chemiluminescent immunosensor was based on immunomagnetic separation and chemiluminescence immunoassay, which utilized a immuno-sandwich scheme as-say with Horseradish peroxidase(HRP) labeled anti-CEA antibody and Immunomagnetic beads(IMBs). And the test instrument consisted of optical detection module and central processing and control system. The optical noise of this instrument was below 40 photons/s. The system could achieve a sample test in 30 minutes, and the detecting result showed a good liner relationship between the intensity of luminescence and concentration of CEA in the range from 1 to1000ng/m L, with a correlation coefficient of 0.9858. Real human samples were also detected with this system in comparison with clinical results and a correlation coefficient of0.9934 was obtained. The instrument is portable and easy to operate, which will provide potential application in cancer clinical test.

关键词： chemiluminescence immunoassay Carcinoembryonic antigen Rapid field test Optical sensor FPGA NIOS II processor

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Paramagnetic particles-based chemiluminescence immunoassay for rapid determination of cardiotrophin-1

Paramagnetic particles-based Chemiluminescence immunoassay f...

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2019年浙江省检验医学学术大会

作者： Ying Ping Zhenping Liu Shufang Qu Yibei Dai Zhihua Tao The Second Affiliated Hospital of Zhejiang University School of Medicine

Objective: Cardiotrophin-1(CT-1), a member of the interleukin-6 superfamily, is a cytokine that can induces hypertrophy and dysfunction in cardiomyocytes. Peripheral blood CT-1 is mainly released from heart via Coronary sinus. Amount studies proposethat CT-1 plasmalevelscanserveas a diagnostic, staging, and prognostic biomarker of cardiovascular diseases. Nevertheless, traditional detection methods of CT-1 are time-consuming and expensive. Theobjectiveofthisstudy is to establish a novel method to detect plasma CT-1 and then evaluated its clinical application for heart failure diagnosis.Design and methods: In this study, we developed a simple paramagnetic particles-based one-step chemiluminescence immunoassay(MPs-CILA) to detect CT-1 in serum/plasma using double antibodies formed sandwich immunoassay scheme. The immunoassay scheme is based on the analyte protein cardiotrophin-1 sandwiched between abiotin-labeled monoclonal antibody(Ab1-Bio) and ananti-CT-1 antibody labelled with acridine ester(Ab2-ACE) to allow luminometer detection. Paramagnetic particles modified with streptavidin(MP-SA) are used in immunoassays to promote the oriented immobilization of the capture antibody(Ab1-SA) on a solid phase. Paramagnetic particles-based assay formathas ahigh magnetic susceptibility,thus realizing rapid and simple separation under external magnets. Conjugatedbiotin-streptavidin system has strong specificity, well stability, ACE has excellent chemiluminescent properties such as low background signal and no need for a catalyst.Luminescence is initiated by oxidation with diluted sodium hydroxide/hydrogen peroxide, the chemiluminescence intensity yield is linearly related to the CT-1 concentration in the standard or sample. Afterwards, 40 heart failurepatients and 20 healthy subjects enrolled as experimental group and control group. The established method was finally applied to detect plasma CT-1 of 40 heart failurepatients and 20 controls, all patients underwent a full clinical evaluation and Cardiac doppler ultrasound.Resul

关键词： Cardiotrophin-1 Paramagnetic particles chemiluminescence immunoassay Acridine ester Determination Heart failure

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Rapid and quantitative analysis of exosomes by a chemiluminescence immunoassay using superparamagnetic iron oxide particles

Rapid and quantitative analysis of exosomes by a chemilumine...

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2019年浙江省检验医学学术大会

作者： Yiyun Wang Zhenping Liu Xuchu Wang Weiwei Liu Zhihua Tao The Second Affiliated Hospital of Zhejiang University School of Medicine

Since the discovery of exosomes, their potential diagnostic value has been the focus of considerable research. However, the lack of a rapid and simple technique for the quantitative analysis of exosomes greatly limits the application of exosomes in clinical research. In this study, we describe a newly developed one-step chemiluminescence immunoassay for the rapid quantitative analysis of exosomes from biofluids. Our new technique, named Exo NANO, adopts a double-antibody sandwich strategy using anti-CD63 antibodyconjugated superparamagnetic iron oxide particles(SIOPs) and acridinium ester(ACE)-labeled anti-CD9 antibodies. SIOPs have narrow size distribution and high magnetic susceptibility, and ACE has excellent chemiluminescent properties such as low background signal and no need for a catalyst. We demonstrated that Exo NANO allows the quantitative analysis of exosomes in the range of 2.92×10 to 2.80×10 particles/μL, with a limit of detection of 2.63×10 particles/μL. Using Exo NANO, we quantified exosomes incell culture medium and clinical biofluids such as serum, saliva, ascitic fluid, and cerebrospinal fluid. We believe that Exo NANO might pave the way for the rapid isolation and quantitative analysis of exosomes for routine clinical applications.

关键词： exosomes superparamagnetic iron oxide particles acridinium ester isolation chemiluminescence immunoassay

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Development of a Magnetic beads chemiluminescence immunoassay for human Neutrophil gelatinase-associated lipocalin

Development of a Magnetic beads chemiluminescence immunoassa...

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2019年浙江省检验医学学术大会

作者： Shufang Qu Ying Ping Danhua Wang Yibei Dai Zhenping Liu Yiyun Wang Zhihua Tao The Second Affiliated Hospital of Zhejiang University School of Medicine Yinzhou No.2 Hospital

Background: Previous studies have shown that blood and urine neutrophil gelatinase-associated lipocalin(NGAL) can be used as a diagnostic indicator for early detection of acute kidney injury(AKI). It is particularly necessary to establish a rapid, sensitive and specificity detection method to detect the levels of NGAL.Methods: A "double-antibody sandwich" strategy was established by using biotin labeled anti-NGAL monoclonal antibody and acridinium ester labeled anti-NGAL monoclonal antibody. Subsequently, magnetic beads coated with streptavidin combined with the double-antibody complex. The concentration of NGAL in serum samples was detected by our chemiluminescence immunoassay and compared with commercial detection method.Results: The limit detection for NGAL concentration of our method is 9.88 ng/ml, and the linear range is 30-3000 ng/ml, required serum sample volume 1.6μl. The intra-CV of the method were 9.40%, 5.63% and 4.55% for three different concentrations of NGAL, and the inter-CV were 14.90%, 14.15% and 11.03% respectively. The recoveries of low, medium and high antigen concentrations were 106.01%, 94.99% and 98.58%, respectively.Conclusions: We constructed a NGAL detection kit with good specificity and high sensitivity. Additionally, this method can be used for automatic simultaneous detection of batch specimens, which is expected to be popularized and applied in automatic detection.

关键词： Neutrophil gelatinase-associated lipocalin Magnetic beads chemiluminescence immunoassay

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A quantitative detection of Cardiotrophin-1 in chronic heart failure by chemiluminescence immunoassay

A quantitative detection of Cardiotrophin-1 in chronic heart...

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2019年浙江省检验医学学术大会

作者： Ying Ping The Second Affiliated Hospital of Zhejiang University School of Medicine

Cardiotrophin-1(CT-1) is a cytokine that could induce cardiomyocytes hypertrophy and dysfunction. Plasma CT-1 might serve as a cardiac biomarker both in diagnosis, staging, and prognostic assessment of heart failure. In this study, a one-step paramagnetic particles-based chemiluminescence immunoassay(MPs-CILA) for rapid and sensitive detection of plasma CT-1 was established. Plasma samples were directly incubated with biotinlabeled anti-CT-1 antibody(bio-Ab) and acridine ester labeled anti-CT-1 antibody(ACE-Ab) to form sandwiched complex. The sandwiched CT-1 was then captured by streptavidin modified paramagnetic particles(MPs-SA) for rapid separation and signal generation. The proposed MPs-CLIA presents a laudable linear relationship ranging from 7.8 pg/mL to 200 ng/mL with a detection limit of 1.0 pg/mL. The recoveries of spiked human plasma samples at low(100 pg/mL), medium(500 pg/mL) and high(800 pg/mL) levels of CT-1 were 104%,108%,110% respectively. The intra-analysis coefficient variation(CVs) of the 3 samples were 6.69%, 3.34% and 3.54%, respectively. And the inter-analysis coefficient variation(CVs) were 10.9%, 3.9% and 4.3% respectively. These results strongly indicate high sensitivity, wide linear range, acceptable precision, and applicable reproducibility of the proposed method to detect plasma level of CT-1. Finally, Plasma CT-1 from 140 subjects with or without chronic heart failure was analyzed to assess the clinical application of MPs-CILA. Noteworthily, the MPs-CLIA method is highly automated such that it is suitable for high-throughput detection of CT-1 in clinical inspection.

关键词： Cardiotrophin-1 Paramagnetic particles chemiluminescence immunoassay Chronic heart failure

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Epidemiological survey of human cytomegalovirus antibody levels in children from Southeastern China

Epidemiological survey of human cytomegalovirus antibody lev...

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2014年浙江省检验医学学术年会

作者：张旗高艳浮苗刘艳清周秋菊余坚郑晓群 School of Laboratory Medicine Wenzhou Medical University

Background This study investigated infection status and distribution of human cytomegalovirus(HCMV)serum markers in hospitalized children from the Wenzhou region.【Methods】This survey was performed on 10,147 hospitalized children from birth to 14 years of age in Southeastern China(Wenzhou region)from March 2010 to March 2013.IgM and IgG antibodies to HCMV were quantitatively detected by chemiluminescence immunoassay(CLIA).HCMV IgM or IgG detection rates,concentration,and distribution in various age groups were retrospectively analyzed.【Results】In this study of hospitalized children,the overall rates of HCMV IgM+and IgG+were 10.8%(1,099/10,147)and 83.0%(8,425/10,147),respectively.The lowest HCMV IgM+rate(1.0%,P0.05).The HCMV IgG+rate was highest in the<28 days group(98.1%,P<0.001).The 28 days months old group had the lowest HCMV specific IgG concentrations(median,133.9 AU/mL,P<0.001).Among 1,099 HCMV IgM+children,405(36.9%)were diagnosed with respiratory infections which pneumonia accounted for 18.2%(200/1,099)of the total population.However,children with respiratory infections had the lowest HCMV IgG concentrations(median,161.1 AU/mL,P<0.05).【Conclusions】HCMV specific antibody responses are very common in hospitalized children with respiratory infection in Wenzhou region.Protection against HCMV airway infection needs greater emphasis and further studies will be helpful to reveal the role of HCMV in children respiratory disease.

关键词： human cytomegalovirus chemiluminescence immunoassay Southeastern China child

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