OBJECTIVETo establish an automatic method for seminal plasma gamma-L-glutamyl transpeptidase (GGT) detection and evaluate its accuracy, repeatability and linear range.

METHODSWe detected the GGT activity in the seminal plasma by rate assay, and established the detection parameters on an automatic biochemical analyzer. Then, we evaluated the reagent blank absorbance, accuracy, repeatability and linear range of the automatic method, and compared the results obtained from the method and the seminal plasma GGT detection kit (Xindi Biological Pharmaceutical Engineering Co., Ltd, Nanjing, China) commonly used in clinical laboratories.

RESULTSThe average absorbance of reagent blank was 0.0476, and the average change rate of blank absorbance (deltaA/min) was 0.000168. The coefficients of variation (CV) for 3 seminal plasma samples with high, middle and low GGT activity detected for 10 times, respectively, were 0.26%, 4.83% and 1.60%. The accuracy of the automatic method was evaluated by a comparison test, and the relative deviation for each concentration point of 40 seminal plasma samples ranged from 13.38% to 11.05%, which met the requirement of < 15%. There was a good linear relationship (r > 0.99) when the seminal plasma GGT activity was between 299 and 1 833 U/L. A significant positive correlation was found between the seminal plasma GGT detection kit (a colorimetric method) as the control and the automatic method as the test reagent in the results of 115 seminal plasma samples (r = 0.981, P < 0.01), with a Kappa value of 0.776 (P < 0.05) and a coincidence rate of 90.43%.

CONCLUSIONThe established automatic method to detect seminal plasma GGT activity has a low reagent blank, good repeatability and accuracy, and fine concordance with the colorimetric method commonly used in clinical laboratories. It is simple, rapid and suitable for screening large numbers of samples, avoids the necessity of diluting the seminal plasma sample, and saves a lot of manpower and reagents.

Automation, Laboratory ; methods ; Humans ; Male ; Reproducibility of Results ; Semen ; enzymology ; gamma-Glutamyltransferase ; analysis

BACKGROUND: Elevated serum alkaline phosphatase (AP) and γ-glutamyl transferase (γ-GT) are commonly observed in patients with acute pyelonephritis. The goal of this study was to examine the clinical significance of elevated serum AP and γ-GT levels and to explore the mechanisms underlying these changes. METHODS: We examined serum AP and γ-GT levels in 438 patients with acute pyelonephritis. Urine AP/creatinine (Cr), urine γ-GT/Cr, fractional excretion of AP, and fractional excretion of γ-GT (FE(γ-GT)) were evaluated in patients with elevated and normal serum levels. AP isoenzymes were also examined. RESULTS: We identified 77 patients (17.6%) with elevated serum AP and 134 patients (30.6%) with elevated serum γ-GT. Among them, both enzymes were elevated in 64 patients (14.6%). Older age, longer hospital stay, elevated baseline serum Cr, and complicated pyelonephritis were associated with increases in serum AP and γ-GT. Multivariate analysis showed that high serum AP levels were significantly correlated with renal impairment (odds ratio, 2.13; 95% confidence interval, 1.08–4.19; P = 0.029). FE(γ-GT) was significantly lower in patients with elevated serum enzyme levels. The liver fraction for AP isoenzyme profile did not increase in patients with elevated serum AP. CONCLUSION: Our results demonstrated that elevated serum AP and γ-GT levels are associated with complicated pyelonephritis and renal impairment. Lower FE(γ-GT) levels in patients with elevated serum enzymes may be the result of decreased urinary excretion of these enzymes.
Alkaline Phosphatase ; gamma-Glutamyltransferase ; Humans ; Isoenzymes ; Length of Stay ; Liver ; Multivariate Analysis ; Pyelonephritis ; Transferases

OBJECTIVETo analyse the factors which influence the four serum fibrosis markers hyaluronic acid (HA), type III procollagen (PCIII), laminin (LN) and type IV collagen (CIV).

METHODSThe levels of serum HA, PCIII, LN and CIV were measured by RIA in 141 patients with chronic hepatitis B (CHB), then the patients were divided into two groups according to the serum fibrosis markers, namely consistent group and inconsistent group. the liver biopsy materials were examined pathomorphologically and liver function was detected by automatic biochemistry analyzer, The interior diameters of the portal vein, the spleen vein and the thickness of the spleen were also measured with ultrasonography.

RESULTS16 patients (14.16%) whose serum fibrosis markers were inconsistent with histological stage of liver fibrosis were found. Their serum fibrosis markers were not correlated with staging of liver fibrosis (P>0.05), but were positively correlated with inflammation grade (x(2)=12.07, P<0.05), at same time, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase(AST), gamma-glutamyltransferase (GGT) and globulin (GLB) decreased obviously, from 89.28 U/L +/- 64.25 U/L to 49.31 U/L +/- 26.75 U/L (t=2.45, P<0.05), 66.10 U/L +/- 42.30 U/L to 40.83 U/L +/- 22.40 U/L (t=2.33, P<0.05), 86.26 U/L +/- 70.36 U/L to 48.99 U/L +/- 29.96 U/L (t=2.08, P<0.05) and 32.13 g/L +/- 5.18 g/L to 28.05 g/L +/- 3.47 g/L (t=3.03, P<0.01) respectively. And the level of albumin (ALB) and the ratio of albumin and globulin (A/G) increased evidently, from 42.34 g/L +/- 4.81 g/L to 46.19 g/L +/- 3.61 g/L (t=3.06, P<0.01) and 1.35 +/- 0.28 to 1.63 +/- 0.26 (t=3.70, P<0.01). But the serum level of alkaline phosphatase (ALP), total bilirubin (TBil), total protein (TP), the width of main portal vein, the width of splenic vein and the thickness of the spleen did not change clearly (P>0.05).

CONCLUSIONAs diagnostic markers, serum fibrosis markers as well as inflammation grade and liver function should be taken into account.

Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Biomarkers ; Female ; Globulins ; analysis ; Humans ; Liver ; physiopathology ; Liver Cirrhosis ; blood ; diagnosis ; physiopathology ; Male ; Middle Aged ; Serum Albumin ; analysis ; gamma-Glutamyltransferase ; blood

OBJECTIVETo evaluate the determination of seminal acid phosphatase (ACP) and gamma-glutamyltranspeptidase (gamma-GT) activity, and analyze the correlation between seminal ACP or gamma-GT and semen parameters.

METHODSACP and gamma-GT activities in 133 samples of seminal plasma were measured. Two of the samples were randomly selected for intra-assay, one for the detection of ACP activity and the other for gamma-GT activity. And another four were selected the same way for the same purpose, two for the detection of ACP activity and the other two for gamma-GT activity. The semen volume, pH, sperm concentration, motility, and grade-a and -b motility were analyzed by CASA system and so were the correlation between seminal ACP or gamma-GT activity and semen parameters.

RESULTSThere was significant positive correlation between ACP and gamma-GT activities (r = 0.570, P = 0.000). The intra-CV of ACP was 13.72%, and inter-CVs of ACP were 13.80% and 15.49%. The intra-CV of gamma-GT was 7.68%, and inter-CVs of gamma-GT were 7.76% and 9.73%. Both seminal ACP and gamma-GT activities had significant negative correlation with pH (r = -0.330, P = 0.000 vs r = - 0. 388, P = 0.000). There was obvious correlation between gamma-GT activity and sperm concentration (r = 0.165, P = 0.045), but not between ACP activity and sperm concentration (r = 0.048, P = 0.546). Neither of seminal ACP and gamma-GT activity was correlated with sperm motility, grade-a and -b motility, semen volume, abstinence time and age.

CONCLUSIONThe precision of the measurement of gamma-GT activity in seminal plasma was higher than that of ACP. The correlation between seminal gamma-GT activity and semen parameters was similar to that between seminal ACP activity and semen parameters. Thus, the determination of gamma-GT activity was a more reliable marker than that of ACP activity for the evaluation of prostate function.

Acid Phosphatase ; analysis ; metabolism ; Adult ; Humans ; Hydrogen-Ion Concentration ; Image Processing, Computer-Assisted ; Male ; Semen ; enzymology ; Sperm Count ; Sperm Motility ; gamma-Glutamyltransferase ; analysis ; metabolism

Human seminal plasma is rich in potential biological markers for male infertility and male reproductive system diseases, which have an application value in the diagnosis and treatment of male infertility. The methods for the detection of semen biochemical markers have been developed from the manual, semi-automatic to the present automatic means. The automatic detection of semen biochemical markers is known for its advantages of simple reagent composition and small amount of reagents for each test, simple setting of parameters, whole automatic procedure with few errors, short detection time contributive to batch detection and reduction of manpower cost, simple calibration and quality control procedure to ensure accurate and reliable results, output of results in the order of the samples in favor of clinical diagnosis and treatment, and open reagents applicable to various automatic biochemistry analyzers. At present, the automatic method is applied in the detection of such semen biochemical markers as seminal plasma total and neutral alpha-glucosidase, acid phosphatase, fructose, &gamma;-glutamyl transpeptidase, zinc, citric acid, uric acid, superoxide dismutase and carnitine, sperm acrosin and lactate dehydrogenase C4, and semen free elastase, which can be used to evaluate the secretory functions of the epididymis, seminal vesicle and prostate, sperm acrosome and energy metabolism function, seminal plasma antioxidative function, and infection or silent infection in the male genital tract.
Acid Phosphatase ; analysis ; Biomarkers ; analysis ; Carnitine ; analysis ; Citric Acid ; analysis ; Epididymis ; metabolism ; Fructose ; analysis ; Humans ; Infertility, Male ; diagnosis ; Isoenzymes ; L-Lactate Dehydrogenase ; Male ; Prostate ; metabolism ; Semen ; chemistry ; Seminal Vesicles ; Spermatozoa ; chemistry ; alpha-Glucosidases ; analysis ; gamma-Glutamyltransferase ; analysis

OBJECTIVETo prepare a monoclonal antibody against hepatoma-specific gamma-glutamyltransferase (GGT-II) and study it's application.

METHODSTwo Bal B/C mice were immunized with pure GGT-II, then their spleen cells were separated and fused to SP 2/0 myeloma cells so as to make hybridoma cell strain which could yield monoclonal antibody against GGT-II. And it's effect of binding GGT-II was detected by competitive inhibitory enzyme linked-immunosorbance assay (ELISA).

RESULTSA mouse hybridoma cell strain which could steadily secrete the monoclonal antibody against GGT-II was obtained and named 2G4F6B2. This monoclonal antibody belonged to IgG1 subclass and was specific to GGT-II, without cross-reaction to GGT-II. The result of detecting human serum GGT-II by ELISA with the monoclonal antibody accorded with that by polyacrylamide gradient electrophoresis.

CONCLUSIONThe monoclonal antibody against GGT-II prepared in this study has high specificity and can be applied in clinic to detect human serum GGT-II.

Animals ; Antibodies, Monoclonal ; Carcinoma, Hepatocellular ; diagnosis ; enzymology ; Enzyme-Linked Immunosorbent Assay ; Female ; Liver Neoplasms ; diagnosis ; enzymology ; Mice ; Mice, Inbred BALB C ; gamma-Glutamyltransferase ; analysis ; immunology

BACKGROUND: Carbohydrate-deficient transferrin (CDT) levels have rarely been determined in an Asian population. We evaluated the analytical performance of a test for measuring CDT levels by using capillary electrophoresis (EP). METHODS: We determined the precision of CDT measurement by using capillary EP and nephelometry and compared the CDT values obtained using both the methods. We included healthy control subjects, abstinent patients with liver disease, and individuals consuming varying amounts of alcohol. RESULTS: The CDT measurement by using capillary EP were correlated well with those CDT measurement by using nephelometry, N Latex CDT assay, Y=0.5706X+1.581, R=0.930. The results obtained from both methods showed good qualitative agreement with each other (kappa coefficient=0.61). Genetic variants of transferrin isoforms were detected in 4.1% of the tested population. Both the CDT and gamma-glutamyl transpeptidase (GGT) levels in the abstinent patients with liver disease were significantly higher than those in healthy abstinent individuals (0.9% vs. 0.5%, 109.5 mg/dL vs. 28.5 mg/dL, respectively), but the difference in CDT values in the 2 groups was less pronounced for the CDT values. Individuals who had a mean daily alcohol intake of more than 60 g/day showed significantly higher CDT levels than those who had a mean daily alcohol intake of less than 60 g/day (1.9% vs. 0.7%, P=0.03). CONCLUSIONS: The CDT test using capillary EP showed good performance, and this method has several advantages such as automation and detection of variant forms. Thus, CDT can be a more useful marker than GGT for monitoring alcohol abstinence, especially in patients with liver disease.
Adolescent ; Adult ; Aged ; Automation ; Electrophoresis, Capillary/*methods ; Female ; Gene Frequency ; Humans ; Liver Diseases, Alcoholic/diagnosis ; Male ; Middle Aged ; Nephelometry and Turbidimetry/methods ; Protein Isoforms/analysis ; ROC Curve ; Republic of Korea ; Transferrin/*analogs & derivatives/analysis ; gamma-Glutamyltransferase/analysis

OBJECTIVETo investigate the value of dynamic examination of duodenal fluid in the differential diagnosis of infantile hepatitis syndrome (IHS) and extrahepatic biliary atresia (EHBA). The aim of the study was to establish a simple, rapid and accurate diagnostic procedure for infantile cholestatic jaundice.

METHODSThe authors developed a special duodenal drainage-tube and established a specific duodenal fluid drainage technique. The duodenal fluids were collected and the colors were documented. The bilirubin, gamma-glutamyltranspeptidase (gamma-GT) and bile acid concentrations in the duodenal fluids were measured.

RESULTSDuodenal fluid drainages were initially performed on 561 cases of infants with cholestatic jaundice. The yellow duodenal fluids were drained within 3-8 minutes after intubation in 342 cases. The yellow fluids were obtained in more patients after continuous drainage for 24 hours (21 cases) and 48-72 hours (16 cases), respectively. The duodenal fluids were light yellowish in 71 cases and white in 111 cases. The drainage techniques were subsequently performed in 182 infants with light yellowish or white duodenal fluids after conservative treatment. The duodenal fluids were yellow in 91 cases, white in 89 cases, and slightly yellowish in 2 cases. The increased levels of bilirubin (> or = 8.5 micromol/L), gamma-GT (> 20 IU/L) and bile acid (positive or 33-260 micromol/L) were observed in the yellow duodenal fluids. While the bilirubin levels were 0-2 micromol/L or 5-8 micromol/L in the white or slightly yellowish duodenal fluids, with gamma-GT levels at 0-5 IU/L and bile acid tested negative. According to the criteria set as bilirubin > or = 8.5 micromol/L, bile acid tested positive and gamma-GT > 20 IU/L in duodenal fluid, 470 infants were diagnosed as HIS; 91 cases were diagnosed as EHBA with duodenal fluid bilirubin < 8.5 micromol/L, bile acid tested negative and gamma-GT < 20 IU/L. The diagnoses of these patients were confirmed by surgical operation.

CONCLUSIONDynamic examination of duodenal fluid is a simple, rapid, safe and reliable method in the differential diagnosis of infantile cholestatic jaundice.

Bile Acids and Salts ; analysis ; Bilirubin ; analysis ; Body Fluids ; chemistry ; Diagnosis, Differential ; Duodenum ; metabolism ; Female ; Humans ; Infant ; Infant, Newborn ; Jaundice, Obstructive ; diagnosis ; Male ; Monitoring, Ambulatory ; instrumentation ; methods ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; gamma-Glutamyltransferase ; analysis

BACKGROUND: Serum Gamma-glutamyltransferase (GGT) has been used clinically as a marker for excessive alcohol consumption or liver diseases, but it was reported recently that GGT is associated with cardiovascular disease. This study was done to verify the association between GGT and the metabolic syndrome in Korean male workers. METHODS: Total cholesterol, triglycerides, HDL cholesterol, body mass index (BMI), waist circumference, blood pressure, fasting glucose, uric acid, and GGT were measured and liver US was performed in 1,215 male workers who underwent annual health check up in a university health promotion center from May to October 2003. The association of GGT with the metabolic syndrome was assessed. RESULTS: The mean age of subjects was 41.9 +/- 7.2 years and the mean BMI was 24.1 +/- 2.7 kg/m2. A raised GGT level (GGT > 75 IU/L) was seen in 172 of 1,136 (15.1%) and the prevalence of the metabolic syndrome was 9.9% (112/1,136). Individuals with the metabolic syndrome had a higher mean GGT concentration (53.4 IU/L) than individuals without the metabolic syndrome (34.4 IU/L; P < 0.001). The subjects with increased GGT showed more risk of metabolic syndrome than the subjects with normal GGT by multivariate analysis (OR=2.835). Subgroup analyses did not change the association between the GGT and the metabolic syndrome. CONCLUSION: This study showed that the serum GGT was associated with the metabolic syndrome and that increased GGT was another feature of the metabolic syndrome.
Alcohol Drinking ; Blood Pressure ; Body Mass Index ; Cardiovascular Diseases ; Cholesterol ; Cholesterol, HDL ; Fasting ; gamma-Glutamyltransferase* ; Glucose ; Health Promotion ; Humans ; Liver ; Liver Diseases ; Male* ; Multivariate Analysis ; Prevalence ; Triglycerides ; Uric Acid ; Waist Circumference

OBJECTIVETo detect serum hepatoma-specific datura stramonium lectin-tightly binding gamma-glutamyl transferase (DSA-GGT) in patients with primary hepatic cancer (PHC) by avidin-biotin ELISA method which was established in our laboratory, and carry on a study of its clinical application.

METHODSTo detect serum DSA-GGT in 45 healthy control subjects, 58 PHC patients and 203 non-PHC patients (including 36 patients with other tumors and 167 patients with benign liver diseases) with the method was established; meanwhile, AFP was detected by ELISA method.

RESULTS38 individuals were DSA-GGT positive in 58 PHC patients, the sensitivity was 65.5%. 18 individuals were DSA-GGT positive in 203 patients without PHC, the specificity was 91.1%. The sensitivity and specificity of AFP in diagnosis of PHC patients was 69.0% and 90.6%, respectively. The sensitivity and specificity of combination of DSA-GGT and AFP was 93.1% and 85.7%, respectively. The average intra-CV and inter-CV of DSA-GGT ELISA was 8.9% and 11.5%, respectively.

CONCLUSIONThe sensitivity and specificity of DSA-GGT ELISA method established in our lab is similar with that of AFP assay and the accuracy is good. Combination of DSA-GGT and AFP may improve the diagnostic sensitivity. The method should be potentially as a new way to improve diagnosis of PHC.

Adult ; Avidin ; Biotin ; Carcinoma, Hepatocellular ; blood ; diagnosis ; enzymology ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Liver Neoplasms ; blood ; diagnosis ; enzymology ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis ; gamma-Glutamyltransferase ; blood