OBJECTIVETo evaluate chemiluminescent immunoassay (CLIA), electrochemiluminescent immunoassay (ECLIA) and ELISA in determination of low level HBsAg.

METHODSAccording to the standard of CLIA Architect i2000, 70 samples were divided into three groups by HBsAg concentration : <1 ng/ml, 1-5 ng/ml and >4 ng/ml. The samples were also determined by ECLIA MODULAR170 and ELISA, and the results were compared with those measured by CLIA Architect i2000.

RESULTSThe concordance rates of ECLIA MODULAR170 with Architect i2000 was 79.2% for <1 ng/ml group, 100% for 1-5 ng/ml group, 100% for >4 ng/ml group. And the concordance rates of ELISA with Architect i2000 was 0 % for <1 ng/ml group, 45.5% for 1-5 ng/ml group and 100 % for >4 ng/ml group.

CONCLUSIONFor determination of low level HBsAg,CLIA Architect i2000 and ECLIA MODULAR170 have high credibility. ELISA is also credible when HBsAg >5 ng/ml, but not for low level HBsAg, especially for HBsAg <1 ng/ml.

Enzyme-Linked Immunosorbent Assay ; Hepatitis B Surface Antigens ; blood ; Humans ; Immunoassay ; methods ; Luminescent Measurements ; methods

We developed a high-sensitivity C-reactive protein quantifiable chemiluminescent immunoassay (hs-CRP CLIA). The high-purity native CRP was purified from hepatic cirrhosis patient ascetic fluid by affinity and ion exchange chromatography and used as an immunogen to develop the monoclonal antibodies (mAbs) against CRP. Twenty-two mAbs were identified reactive with CRP in ELISA and 13 of them were reactive in the phosphorycholine ligand capture ELISA. The mAbs 10C5 and 10C11 were selected to develop the hs-CRP CLIA. The linearity and performance of the hs-CRP CLIA was characterized. It was showed not reactive when testing against other serum materials (IgG, hemoglobin and triglyceride). The reliable correlation (R2 > 0.993) was obtained between testing value (RLU/S) and the concentration of human serum CRP calibrator. The linearity fell in the range of 0.04-20.38 mg/L. The assay has good accuracy and reproducibility, the mean recovery was 99% and the precision of the intra- and inter assay was CVs (4.2%-5.8%) and (9.0%-11.5%), respectively. In testing of 90 human sera, this assay performed well and correlated comparably with a commercial hs-CRP ELISA kit. Thus, hs-CRP CLIA is an accurate, reliable, quantifiable assay for detection of high-sensitive C-reactive protein in serum, it may be useful to improve the risk assessment of cardiovascular disease and the prognosis of inflammatory bowel disease.
C-Reactive Protein ; analysis ; chemistry ; Humans ; Immunoassay ; methods ; Luminescent Measurements ; methods ; Sensitivity and Specificity

Chromatography, High Pressure Liquid ; methods ; Drug Residues ; analysis ; Electrochemistry ; Ethidium ; analysis ; Luminescent Measurements ; methods ; Sensitivity and Specificity

AIMTo study the sensitizing effect of acemetacin (ACE) on the weak chemiluminescent (CL) reaction of KMnO4 with sulfite and establish a fast and convenient method for CL detection of ACE.

METHODSUsing the sensitizing effect of ACE on KMnO4-Na2SO3 system and flow injection technique to determine the concentration of ACE.

RESULTSUnder optimal conditions, the CL intensity of 1.0 x 10(-2) mol x L(-1) H3PO4 - 5.0 x 10(-5) mol x L(-1) KMnO4 - 4.0 x 10(-4) mol x L(-1) Na2SO3 was proportional to the concentration of ACE ranging from 1.0 x 10(-7) to 1.0 x 10(-5) mol x L(-1). The detection limit of ACE was 6.9 x 10(-8) mol x L(-1) at 3sigma. Satisfactory results were obtained for determination of ACE at 2.5 x 10(-6) mol x L(-1).

CONCLUSIONThe present method showed good precision, high sensitivity and selectivity and could be used for fast and convenient detection of ACE. It would be of significance to the clinical and pharmacological study of acemetacin.

Flow Injection Analysis ; methods ; Indomethacin ; analogs & derivatives ; analysis ; Luminescent Measurements ; methods ; Potassium Permanganate ; chemistry ; Sulfites ; chemistry

OBJECTIVETo perform a systematic comparative analysis of two different commercial automated systems using chemiluminescence immunoassay to quantitatively detect hepatitis B virus surface antigen (HBsAg) in patient sera.

METHODSThe Elecsys2010 electrical chemiluminescence immunoassay (ECLIA; manufactured by Roche) and the ARCHITECT il000 chemiluminescence magnetic microparticle immunoassay (CMIA; manufactured by Abbott) were used to detect HBsAg in 100 serum samples of individuals who presented at our department with suspected hepatitis infection between January and May 2012. The manufacturer's protocols were strictly followed. The categorical data was analyzed by Chi-squared test, and linear regression analysis was used to compare the results of the two assay systems.

RESULTSThe HBsAg detection results from the two different assay systems showed good correlation (r >or= 0.95), and had good correlation at a low (r = 0.966), medium (r = 0.974) and high (r = 0.984) cutoff values. However, the positive detection rate of CMIA was significantly higher than that of ECLIA(94% vs. 88%, P < 0.05). When the HBsAg content was below 0.10 IU/ml, the ECLIA detection rate and sensitivity were slightly higher than those of CMIA.

CONCLUSIONThe ARCHITECT i1000 and Elecsys 2010 immunoassay systems have good correlation in quantitative detection of HBsAg, but the former may be more sensitive.

Hepatitis B Surface Antigens ; blood ; Humans ; Immunoassay ; methods ; Luminescent Measurements ; Sensitivity and Specificity

Animals ; Antibodies, Bacterial ; blood ; Luminescent Measurements ; methods ; Streptococcus suis ; immunology ; Swine

AIMTo establish a simple and novel method for the determination of vitamin B2 rapidly in pharmaceutical preparations.

METHODSVitamin B2 was determined by a chemiluminescence (CL) sensor combined with flow-injection (FI) technology. The analytical reagents involved in the CL reaction, luminol and hexacyanoferrate (III), were both immobilized on an anion-exchange resin column in FI system. The CL signal produced by the reaction between luminol and hexacyanoferrate (III), which were eluted from the column through sodium phosphate injection, decreased in the presence of vitamin B2.

RESULTSThe decreased CL intensity was linearly correlated with the vitamin B2 concentration in the range of 0.01-1.0 microgram.mL-1, the detection limit was 4.0 ng.mL-1 vitamin B2 (3 sigma). At a flow rate of 2.0 mL.min-1, the procedure including sampling and washing could be performed in 2 min with a relative standard deviation of less than 3.0%.

CONCLUSIONThe flow sensor exhibited both good sensitivity and stability. It could be reused more than 450 times and has been applied successfully to the analysis of vitamin B2 in pharmaceutical preparations.

Flow Injection Analysis ; methods ; Luminescent Measurements ; Luminol ; Riboflavin ; analysis ; Tablets ; chemistry

We use direct-write laser micromachining technology to fabricate the microfluidic chip, and to establish a microfluidic chemiluminescence immunoassay system based on superparamagnetic microbeads, for detecting alpha- fetoprotein (AFP). The AFP analysis can be completed in 20 minutes with 5 microl sample and 5 microl reagent, and there is a good linear correlation in the range of 1-800 ng/ml.
Equipment Design ; Humans ; Immunoassay ; instrumentation ; methods ; Luminescent Measurements ; instrumentation ; methods ; Microfluidic Analytical Techniques ; instrumentation ; alpha-Fetoproteins ; analysis

Fluorescence ; Host-Pathogen Interactions ; Luminescent Measurements ; methods ; Luminescent Proteins ; chemistry ; genetics ; metabolism ; Plant Diseases ; virology ; Plant Viruses ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; genetics ; metabolism ; virology

Quantitative specific detection of Staphylococcus aureus is based on recombinant lysostaphin and ATP bioluminescence. To produce recombinant lysostaphin, the lysostaphin gene was chemically synthesized and inserted it into prokaryotic expression vector pQE30, and the resulting expression plasmid pQE30-Lys was transformed into E. coli M15 for expressing lysostaphin with IPTG induction. The recombinant protein was purified by Ni(2+)-NTA affinity chromatography. Staphylococcus aureus was detected by the recombinant lysostaphin with ATP bioluminescence, and plate count method. The results of the two methods were compared. The recombinant lysostaphin was successfully expressed, and a method of quantitative specific detection of S. aureus has been established, which showed a significant linear correlation with the colony counting. The detection method developed has good perspective to quantify S. aureus.
Adenosine Triphosphate ; chemistry ; Chromatography, Affinity ; Escherichia coli ; Luminescent Measurements ; methods ; Lysostaphin ; chemistry ; Recombinant Proteins ; chemistry ; Staphylococcus aureus ; isolation & purification