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We aim to establish a chip-based digital PCR (dPCR) method for detecting copy number variation of the LAPTM4B gene in non-small cell lung cancer (NSCLC), and preliminarily evaluate its basic performance and clinical feasibility. The LAPTM4B gene primers and specific probes were designed to establish a dPCR reaction system. The detection limit, precision, and linearity of the method were verified according to the prepared target DNA samples of different concentrations. The reaction system of dPCR for LAPTM4B gene copy number detection was established and optimized for the first time. The results showed that 12. 5% of LAPTM4B gene copy number deletion could be detected at the lowest level. The coefficient of variation of inter-batch precision was less than 10%, and the linearity of deletion ratio was good in the range of 12. 5%-100% (R
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<p><b>OBJECTIVE</b>To understand the incidence of acute kidney injury (AKI) in infants and toddlers and evaluate the possibility of predicting AKI with urine neutrophil gelatinase-associated lipocalin (NGAL), interleukin 18 (IL-18), N-acetyl-beta-D-glucosaminidase (NAG), microalbumin (MA) and α1-microglobulin (α1-MG) after surgeries for congenital heart diseases with cardiopulmonary bypass (CPB).</p><p><b>METHOD</b>Fifty-eight children (ages ≤ 3 years) who had undergone surgery for congenital heart diseases with CPB were enrolled. Urinary samples were collected before and 4 h, 6 h, 12 h, 24 h post CPB to detect the concentration of NGAL, IL-18, NAG, MA and α1-MG.</p><p><b>RESULT</b>The AKI group had 29 cases, none AKI group also had 29 cases. Urinary concentration of NGAL 4, 6, and 12 h post CPB were significantly higher in AKI group (2820 µg/g, 905.7 µg/g, 76.1 µg/g separately) than in none AKI group (27.6 µg/g, 19.5 µg/g, 16.0 µg/g separately, P < 0.01). Urinary concentration of IL-18 4, 6, 12 and 24 h post CPB were significantly higher in AKI group than in none AKI group (P < 0.05). Urinary concentration of NAG 4 h and 6 h post CPB were significantly higher in AKI group than in none AKI group (P < 0.01). Urinary concentration of MA/UCr post CPB 4 h, 6 h and 12 h were significantly higher in AKI group than in none AKI group (P < 0.05). Urinary concentration of α1-MG/UCr post CPB 4 h, 6 h and 12 h were significantly higher in AKI group than in none AKI group (P < 0.01). All the five biomarkers had predictive abilities at 4-hour after surgery.</p><p><b>CONCLUSION</b>Urine biomarkers NGAL, IL-18, NAG, MA and α1-MG were valuable early predictors of AKI after CPB surgery.</p>
الموضوعات
Child, Preschool , Female , Humans , Infant , Male , Acute Kidney Injury , Urine , Acute-Phase Proteins , Urine , Alpha-Globulins , Urine , Biomarkers , Urine , Cardiopulmonary Bypass , Creatinine , Urine , Heart Defects, Congenital , General Surgery , Interleukin-18 , Urine , Lipocalin-2 , Lipocalins , Urine , Predictive Value of Tests , Proto-Oncogene Proteins , Urine , Sensitivity and Specificityالملخص
<p><b>OBJECTIVE</b>To explore the therapeutic efficacy of a combined treatment modality using transcatheter arterial chemoembolization (TACE) and percutaneous ethanol injection (PEI) to treat hepatocellular carcinoma (HCC) complicated with main branch intraportal vein tumor thrombosis (PVTT).</p><p><b>METHODS</b>Clinical data was collected retrospectively for patients diagnosed with and treated for HCC plus main branch PVTT at our hospital between January 2007 and January 2010. The total study population (n = 51) consisted of 38 males and 13 females, with an average of 50.1 years (range: 24-73). Among these patients, 26 had been treated with TACE + PEI (group A) and 25 had been treated with TACE alone (group B). Short-term changes in PVTT (i.e. disappearance, shrinkage, and/or stability) and tumor (i.e. complete response, partial response, and/or stable disease) were assessed by using the t-test (continuous variables) or the Chi-squared or Fisher's exact tests (categorical variables). Between-group differences in survival time were assessed by the Kaplan-Meier analysis and log-rank test.</p><p><b>RESULTS</b>The follow-up time ranged from 3-24 months after treatment, and no serious treatment-related complications were recorded for any of the patients (0/51). The time of TACE treatment was significantly longer for the patients receiving the combination therapy (group A: 3.21.4 vs. group B: 2.40.9, t = 2.22, P = 0.032). The patients in group A received between 2-8 PEI treatments. The TACE + PEI combined treatment showed significantly better therapeutic efficacy for PVTT (group A: 19/26 vs. group B: 10/25, X2 = 5.685, P = 0.019). The tumor response was significantly better in patients treated with TACE + PEI at post-treatment month 3 (group A: 20/26 vs. group B: 18/25, X2 = 0.163, P = 0.705) and month 6 (group A: 17/20 vs. 10/19, X2 = 2.58, P = 0.027). Finally, the average survival time was significantly better in patients treated with TACE + PEI (group A: 12.856.02 months (range: 5-23) vs. group B: 8.653.39 months (range: 4-16), t = 3.051, P = 0.004).</p><p><b>CONCLUSION</b>TACE + PEI combination therapy for main branch PVTT in HCC patients is more efficacious than TACE alone, and is associated with a longer survival time.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Pathology , Therapeutics , Chemoembolization, Therapeutic , Combined Modality Therapy , Ethanol , Injections, Subcutaneous , Liver Neoplasms , Pathology , Therapeutics , Neoplastic Cells, Circulating , Portal Vein , Pathology , Retrospective Studies , Thrombosisالملخص
<p><b>BACKGROUND</b>The equations for estimating glomerular filtration rate (GFR) based on creatinine have been found to have limitations and have not been generalizable across all populations. Equations based on cystatin C provide an alternative method to estimate GFR. Whether the equation based on cystatin C alone or combined creatinine would improve GFR estimates has not been validated among Chinese patients with chronic kidney disease (CKD) and diabetes. The aim of this study was to compare the performance of the modification of diet in renal disease (MDRD) equation based on creatinine with the five cystatin C-based formulae for estimation of GFR in patients with CKD and diabetes.</p><p><b>METHODS</b>A total of 166 patients with CKD and 91 patients with type 2 diabetes were enrolled in this study. Cystatin C was measured by using the particle-enhanced immunonephelometric method and estimated formulae proposed by five different investigator teams (Stevens, Ma, Rule, Macisaac and Perkins). The plasma clearance of (99m)Tc-DTPA was determined as measured GFR (mGFR).</p><p><b>RESULTS</b>For CKD patients, the bias and accuracy for the Ma and Macisaac equations were superior compared with the MDRD, and the mean results for the Ma formula were closer to mGFR than the other equations in CKD stages 2 - 5. The differences between Macisaac and mGFR in CKD stages 2 - 4 were significantly less than those in CKD stage 1 or 5. Stevens and Rule's formulae revealed a similar bias and accuracy compared with the MDRD equation. The MDRD formula had a higher accuracy in CKD stages 3 - 5 as compared with the results in other stages. For diabetic patients, the mean results between Macisaac and mGFR were closer than those of other equations in mGFR >or= 90 mlxmin(-1)x1.73 m(-2) stage. In GFR 60 - 89 mlxmin(-1)x1.73 m(-2) stage, the MDRD formula showed the smallest difference compared with other equations. All equations overestimated GFR in the cases with GFR < 60 mlxmin(-1)x1.73 m(-2) stages. The MDRD formula had a greater accuracy within 50% of mGFR than the equations based on cystatin C in diabetic patients. Perkins formula showed a large positive bias and low accuracy, therefore it may not be suitable for assessing GFR in patients with CKD and diabetes.</p><p><b>CONCLUSIONS</b>The formulae for estimating GFR based on cystatin C or creatinine have different trends and accuracies in patients with CKD and diabetes, especially in patients with various GFR levels. The equations based on cystatin C provide less accurate results than MDRD formulae, at least in the diabetic patients. Therefore, whether the formulae based on cystatin C are superior to MDRD formula requires further investigation in large diverse populations.</p>
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Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Chronic Disease , Creatinine , Blood , Cystatin C , Blood , Diabetes Mellitus , Glomerular Filtration Rate , Kidney Diseasesالملخص
<p><b>BACKGROUND</b>Serum creatinine (Scr) measurement plays a key role in glomerular filtration rate estimation (eGFR), chronic kidney disease (CKD) diagnosis as well as CKD treatment. However, the test results of Scr from different laboratories vary significantly. In order to get comparable results, the European in vitro diagnostic (IVD) directive requires traceability to reference methods and materials. The purpose of this study was to verify the effect of traceability implementation by investigating the trueness of creatinine measurement on nine homogenous systems in Beijing.</p><p><b>METHODS</b>Commutable frozen human serum reference material, National Institute of Standards & Technology (NIST) Standard Reference Material (SRM) 967, was used to verify the trueness of Scr measurement results from nine homogeneous analytical systems of seven companies which are the most widely used systems in Beijing's third-grade hospitals. The methods referred to the Jaffe's and Enzymatic methods.</p><p><b>RESULTS</b>from nine routine measurement systems were assessed using two criteria: biological variability and Clinical Laboratory Improvement Amendments' 88 (CLIA' 88). We simulated a series of broken lines representing the limits of SD and bias that would produce a relative increase (or decrease) of 10% and 20% in the measurement error when estimating GFR (MEeGFR) using the isotope dilution mass spectrometry (IDMS)-traceable Modification of Diet in Renal Disease (MDRD) Study equation.</p><p><b>RESULTS</b>of the College of American Pathologists (CAP) 2008-B LN24 Survey were compared with our investigation results.</p><p><b>RESULTS</b>Compared with the total error criteria of biological variability, Ortho (traceable to IDMS) met the minimum acceptable criteria; Roche (Jaffe), Roche (Enzymatic), Shino and Daiichi met the desirable criteria at level I. At level II, Ortho (traceable to gas chromatography/isotope dilution mass spectrometry, GC/IDMS), Dade Behring and Beckman (traceable to rate Jaffe) met the minimum acceptable criteria; Roche (Enzymatic) met the optimum criteria. The other five systems met the desirable criteria. Compared with the second criterion, all the results met the requirement of CLIA' 88. Trueness evaluation showed: the MEeGFR of Dade Behring exceeded 10% while the MEeGFRs of Beckman (traceable to rate Jaffe), Beckman (traceable to IDMS) and Ortho (traceable to Jaffe/High Performance Liquid Chromatography) exceeded 20% at level I. At level II the MEeGFRs of Dade Behring, Ortho (traceable to GC/IDMS) and Beckman (traceable to rate Jaffe) exceeded 10%. None of the nine systems got a MEeGFR higher than 20%. The conclusions of NIST SRM 967 agreed with those of LN 24 except for the Beckman measurement system.</p><p><b>CONCLUSIONS</b>Trueness investigation of routine creatinine assays on nine homogeneous systems demonstrates an encouraging outcome that meets clinical requirements. Among the nine homogeneous routine systems, Roche and Daiichi produce the most accurate results. The implementation of traceability is effective.</p>
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Humans , Creatinine , Blood , Gas Chromatography-Mass Spectrometry , Glomerular Filtration Rateالملخص
<p><b>OBJECTIVE</b>To determine serum carnitine levels in patients with liver diseases and to investigate their significance.</p><p><b>METHODS</b>25 patients with acute viral hepatitis, 34 with chronic viral hepatitis, 22 with post hepatitis cirrhosis with normal renal function, 9 with post hepatitis cirrhosis but with renal disfunction, and 40 healthy subjects (serving as controls) were enrolled in this study. An enzymatic cycling method was used to determine the serum free carnitine levels.</p><p><b>RESULTS</b>The serum free carnitine level was (48.3+/-10.2)micromol/L in the healthy control group. It was (35.2+/-13.2)micromol/L in the acute viral hepatitis group, (36.5+/-9.9)micromol/L in the chronic viral hepatitis group, (45.0+/-11.0)micromol/L in the post hepatitis cirrhosis with normal renal function group, and (83.6+/-50.4)micromol/L in the post hepatitis cirrhosis with renal dysfunction group. Serum free carnitine levels in the acute viral hepatitis and chronic viral hepatitis groups were significantly lower than those in the healthy controls. There were no significant differences in serum free carnitine levels of the post hepatitis cirrhosis group and the normal control group.</p><p><b>CONCLUSIONS</b>Patients with liver diseases can have carnitine metabolism errors. One of the secondary carnitine lack causes is liver disease.</p>
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Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carnitine , Blood , Chronic Disease , Hepatitis, Viral, Human , Blood , Liver Cirrhosis , Bloodالملخص
Objective To compare application in Beijing population using the diagnostic criterias for metabolic syndrome(MS)according to the criterias of the International Diabetes Federation(IDF),Third Report of the National Cholesterol Education Program(NCEP)Expert Panel on Detection,Evaluation,and Treatment of High Blood Cholesterol in Adults[ATP Ⅲ(revised)]and Chinese Diabetes Society(CDS), respectively.Methods This study was based on an investigation on the risk factors of cardiovascular diseases involving 4 628 cases.MS was diagnosed according to these three definitions respectively and prevalence derived from these three definitions was compared.Results The prevalence of MS were 40.3 %, 44.3% and 35.7% according to the IDF,ATP Ⅲ(revised)and CDS respectively.The agreement in the diagnosis of MS using IDF and CDS,ATP Ⅲ(revised)and CDS,IDF and ATP Ⅲ(revised)were 81.9%, 83.7% and 96.0% respectively.There were 4.0% subjects among the MS patients diagnosed by ATP Ⅲ (revised)who can not be screened by IDF criteria.The prevalence of obesity was 19.6% according to ATP Ⅲ,much lower than the prevalence using CDS and IDF,whereas this rate increased to 58.6% using ATP Ⅲ(revised)criteria,showing diagnostic agreement with CDS.The coincidence rate between ATP Ⅲ (revised)and CDS was 80.6% Conclusion This investigation shows good agreement in the diagnosis of MS using IDF,CDS and ATP Ⅲ(revised)among Beijing population and these criterias can be used in China.
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Objective To evaluate the value of serum creatinine(Scr),creatinine clearance (Ccr),MDRD equation 7(MDRD 7),~(99)Tc~m-DTPA renal dynamic imaging(gGFR)and cystatin C in screening changed glomerular filtration function in type 2 diabetic patients.Methods The ~(99)Tc~m-Diethylene Triamine Pentaacetic Acid(~(99)Tc~m-DTPA)plasma clearance(rGFR)obtained with the dual plasma sampling method was used as a reference with which Scr,Ccr,MDRD 7,gGFR and Cystatin C were compared.Results Sixty of type 2 diabetic patients were selected,including 35 male and 25 female.The average age was(62.4?11.7)years and the average diabetic history was(10.66?9.35)years.Scr,Ccr, MDRD 7,gGFR were all correlated significantly with rGFR.Correlation coefficients were 0.675 for Ccr, -0.588 for Cystatin C,-0.500 for Scr,O.428 for MDRD 7,0.367 for gGFR(P values all
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Objective Understand the variation of serum creatinine measurement in clinical laboratories of some hospitals in Beijing.Methods 8 samples of mixed frozen human serum added different creatinine concentration standard materials(the creatinine concentration were80-1 000 ?mol/L)and 8 samples of mixed frozen serum of patients(contained different creatinine concentration)were distributed to 13 clinical laboratories(31 series of detection systems)with the way of spot investigation.Every clinical laboratories measured the samples followed the standard operating procedure.Results As to the mixed frozen human serum added different creatinine standard materials,the CV of different detection systems results were 5.74%-9.68%;as to the mixed frozen patients' serum,the CV was 5.90%-11.69%. Compared with Beckman closed detection systems,the results of Dade systems(which used the kinetic alkaline pieric acid method)showed the bias were-5.99%-0.35%,and as to the other systems which measured by alkaline picric acid method,when creatinine concentrations were 200 ?mol/L,the results showed negative bias,and the greatest bias was-8.45%.The bias plots revealed negative for all of the detection systems with enzymatic method over the whole concentration range,and the greatest bias was -8.88%.Conclusions The creatinine determination results of Beckman and Dade closed detection systems were consistent.The results of detection systems which used enzymatic method were generally lower than Beckman detection systems.What's more,the creatinine measurement variations of clinical laboratories were very large,especially for the results of unclosed detection systems,so it was urgent need to solve the standardization of creatinine measurement.
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Objective To evaluate the performance of Cys C results among two detection system.Methods The particle-enhanced immunonephrometic assay was used in Dade Behring BNII. Immunoturbic assay was used in Hitachi 7170 to evaluate the JING' YUAN reagents.We compared the precison,linearity,interference,correlation,and calibrators agreement with Dade Behring BNII.Results The total CV of the samples that contain 0.6-5.0 mg/L was less than 10%.The Dade Behring and JING'YUAN method showed good linearity.Haemoglobin(10 g/L),Bilirubin(300 mg/L), Vitamin C(5 g/L)in the tested sample had no significant interference in the assay(interference 0.05) between JING' YUAN and Dade Behring reagents.Values were slightly lower than that from the Dade Behring BNII method,the mean bias was-0.16.The bias range was 1.1%-23% between JING'YUAN and Dade Behring for one sample.Conclusions The precision,linearity and interference test were suitable for routine Cys C measurement on automated biochemistry analyzer,but results has bias.
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Chronic kidney disease(CKD)is a major public health problem worldwide. Understanding by doctors and laboratorians of the importance of reliable serum creatinine measurement in GFR estimation and of factors that may affect creatinine measurement is critical to ongoing public health efforts to improve the diagnosis and treatment of patients with CKD.We present an overview of the commonly used methods,their performances and limitations and the required performance criteria for the measurement of serum creatinine.Available resources for standardization of serum creatinine measurements and recommendations for creatinine measurement and GFR estimations are introduced.
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To explore the possibility of a new therapeutic strategy for leukemia by intervening in the DNA methylation to re-express p15 suppressor gene, methylation inhibitors, 5-Aza-2'-deoxycytidine (5-Aza-CdR) and cell differentiation agent (CDAII) were used to treat myelogenous leukemia cell line KG1a in which p15 gene expression was suppressed due to DNA hypermethylation. The biological characteristics of KG1a cells untreated or treated with the agents were investigated and analyzed using morphology, methylation specific-PCR (MSP), (3)H-labeled microassay technique, restriction endonuclease reaction, flow cytometry and immunofluorescence methods. The results indicated that both agents showed concentration-dependent and time-dependent inhibition of cell proliferation. 5-Aza-CdR and CDAII induced apoptosis and cell differentiation with G(2) and G(0)/G(1) arrest respectively. Furthermore, DNA methyltransferase activity and level of methylation in genomic DNA were decreased and p15 protein was re-expressed partially. It is concluded that it is possible to treat leukemia by intervening in the DNA methylation using methyltransferase inhibitors and it is worth to make a thorough study on mechanism of the new strategy.
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Humans , Azacitidine , Pharmacology , Cell Cycle Proteins , Genetics , Cell Differentiation , Cell Division , Cell Line , Cyclin-Dependent Kinase Inhibitor p15 , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , DNA Methylation , DNA Modification Methylases , Dose-Response Relationship, Drug , Enzyme Inhibitors , Pharmacology , Genes, Tumor Suppressor , Leukemia, Myeloid , Drug Therapy , Pathology , Tumor Suppressor Proteinsالملخص
Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.
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Objective To compare the coherence of serum creatinine,creatinine clearance(Ccr), Cystatin C,and estimated glomerular filtration rate(eGFR)in each stage of chronic kidney disease(CKD) patients.Methods Creatinine in serum and urine were determined by Jaffe method;serum Cystatin C was measured by particle enhanced turbidimetric method,while eGFR was calculated using the abbreviated Modification of Diet in Renal Disease(MDRD)equation which was mainly based on the serum creatinine concentration.According to the American national kidney foundation-Kidney Disease Outcome Quality Initiative(NKF-K/DOQI)guideline,all cases were grouped by eGFR into 5 stages.Results In these 228 cases,as eGFR decreased gradually,the average levels of creatinine and Cystatin C increased,while Ccr decreased.The level of each items showed a statistic difference among each stage(P0.05);in eGFR 60-89 ml/min group,the average level of creatinine was 83.3 ?mol/L,the abnormal rate was only 6.8%,it was not a sensitive marker to detect the slightly damaged GFR,the levels of Ccr and Cystatin C showed a marginal decrease and increase,with an abnormal rates of 70% and 86%,there was a statistic difference among the three abnormal rates(P
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Objective To evaluate the sensitivity and specificity of new genevation Beckman Coulter cTnI in diagnosis of non-ST-elevation myocardial infarction(NSTEMI),and the values of cTnI in prognosis of non-ST elevation acute coronary syndromes(NSTEACS).Methods The cTnI was determined in NSTEMI patients and in control group,then analyzed the data with receiver operating characteristic curve (ROC curve)statistical software.Two hundred and sixty-nine patients with non-ST elevation acute coronary syndromes were divided into cTnI≥0.04 ?g/L and cTnI
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Objective To investigate the accuracy and comparability of ?-glutamyltransferase (?- GT) measurement results on human serum samples and controI materials before and after calibration with a common human serum calibrator.Methods A human serum calibrator was prepared by pooling fresh serum aliquots and assigning a value for ?-GT with the IFCC reference method.The calibrator together with 5 human serum samples and 10 control samples were sent to 15 clinical laboratories and the sermn and control samples were measured with different analytical systems before and after a calibration with the calibrator.The results were analyzed for biases and interlaboratory variations.Results For the serum samples,the calibration resulted in reductions in biases from -9.0%~-14.2% to -0.8%~-7.9%,and in interlaboratory variations from 6.9%~11.6% to 2.8%~4.4%.No improvement was observed on the control samples.Conclusions Accuracy and comparability of serum ?-GT measurements can be improved by using a common human serum calibrator.Some control materials may not be commutable for human serum in ?-GT measurements.
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98.95%.The results of certified reference materials were consistent with the target value,and average deviation was -0.31%~1.35%.HPLC was served as the independent variable.When ereatinine was 200 ?mol/L,Bias of Beckman LX20 system,Vitros dry chemistry system and enzymatic method were -10%~13%,-13%~14% and -20%~10%,respectively.Bias of enzymatic method results was mostly negative,when creatinine was
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Objective To investigate the efficacy of using a calibrator with values assigned with the reference method for improving the comparability of serum gamma-glutamyltransferase (GGT) measurements.Methods The IFCC reference method for GGT was established and the performance was verified by testing a certified reference material (CRM).A calibrator was prepared and its value for GGT was assigned with the reference method.Forty serum samples were measured on different (including HITACHI 7600,7060,7170,7180 and BECKMAN LX20,OLYMPUS AU 400) chemistry analyzers with Zhongsheng GGT reagent kits calibrated with the calibrator.The samples were also measured on the same analyzers using a theoretical factor.Biases of results obtained with the calibration and with the theoretical factor based calculation were compared.Results The reference method resuhs on the CRM agreed the certified value within the stated uncertainty.Serum results calculated from the theoretical factor showed various biases and inter-analyzer variations.When the analyzers were calibrated with the calibrator,the number of results with biases less than 10% became significantly higher and those with biases more than 20% significantly lower.The variation of the results on 5 serum samples was reduced from 11.0%~14.0% to less than 5% by using the calibrator.Conclusion Accuracy and comparability of GGT measurements with of ZhongSheng GGT kits can be improved by using a calibrator that has a value assigned with the reference method.
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80 mL/ min were 12 cases;minor lesion group(50