Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:APRD00691 (EE2)
 7,802 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An enzymatic rate assay is described for measuring cholesterol in serum. Cholesterol is analyzed by mixing 5 mul of sample with a reagent consisting of cholesterol esterase, cholesterol oxidase, catalase, acetylacetone, methanol, and hydroxypolyethoxydodecane in a ammonium phosphate buffer at pH 7.0. The rate of increase in absorbance of the dihydrolutidine product is measured at 37 degrees C and 405 nm. The change in absorbance between 4 and 10 min is used to calculate the cholesterol concentrations by using simultaneously determined free cholesterol standards. The change is linearly related to cholesterol concentration up to 4 g/liter. Samples containing bilirubin up to 200 mg/liter, uric acid up to 200 mg/liter, and hemoglobin up to 1 g/liter, or certain drugs (clofibrate, phenobarbital, nicotinic acid, salicylate, Ketochol, and Ovral) gave no interference. Ascorbic acid added to serum caused a positive interference. Lipemic samples gave values that were slightly lower than did the method of Abell et al., used for comparison. Our kinetic assay, compared with the method of Abell et al., the enzymatic assay used with Abbott's Bichromatic Analyzer, and the Technicon SMA 12/60 enzymatic procedure gave correlation coefficients of 0.992, 0.985, and 0.986, respectively.
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PMID:Enzymatic rate method for measuring cholesterol in serum. 1 1

We describe the effects of uric acid, hemolysis, drugs, ascorbic acid, lipemia, and bilirubin on the enzymic measurement of cholesterol in serum by use of reagent kits from Abbott, Beckman, Boehringer Mannheim, Calbiochem, and Worthington. In all of these, the chromogen formed from the reaction of hydrogen peroxide with phenol and 4-aminoantipyrene is measured. The absorbance was measured at 500 nm vs. a serum blank for each kit--except Abbott's with which the recorded absorbances were the differences between readings at 500 and 600 nm. With all reagents kits, there was no interference from uric acid up to 200 mg/liter, hemoglobin up to 1.0 g/liter, or drugs (clofibrate, phenobarbital, Ketochol, Ovral-28), but negative interferences from ascorbic acid. Except for the Abbott kit, the cholesterol values obtained for lipemic samples were lower than found with the comparison method [Abell et al., Stand. Methods Clin. Chem. 2, 26 (1958)]. With Abbott's reagents, for most lipemic samples, the values were the same. Bilirubin at concentrations of 200 mg/liter significantly decreased the cholesterol values with Beckman, Calbiochem, and Worthington reagent kits. With Boehringer Mannheim reagent a small negative interference was observed and with Abbott reagent a small positive interference was observed when the bilirubin concentrations were 200 mg/liter.
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PMID:Interference with the enzymic measurement of cholesterol in serum by use of five reagent kits. 84 75

One first generation assay (manufactured by Ortho, test I) and 3 second generation anti-HCV ELISAs (manufactured by Ortho, Abbott, and UBI, tests II-IV) were compared. Sera from 4 different sources were used: (1) intravenous drug-users (IVDUs, n = 50), (2) blood donors (n = 1055), (3) all clinical samples from one day of routine anti-HCV testing (n = 89), (4) hemodialysis patients previously found negative by test I but clinically suspected to have a HCV infection (n = 11). Confirmatory anti-HCV tests were carried out with a second generation recombinant immunoblot assay (RIBA II). In sera positive exclusively by test IV, one antibody consumption test (UBI HCV Neutralization EIA) and one further immunoblot assay (INNO-LIA HCV Ab) were used. PCR for HCV RNA was carried out on all hemodialysis patient sera and in the RIBA II positive blood donor sera. The second generation ELISAs discriminated 11 more positive samples than the first generation test (2 IVDUs, 5 blood donors, 4 clinical samples). The 9 sera from blood donors and clinical samples were all RIBA II positive or indeterminate. The second generation tests thus showed increased sensitivity. The second generation tests also showed increased specificity in that 4 samples that were positive by test I but negative by the second generation tests, were also negative by RIBA II. With few exceptions, all RIBA II-positive and most of the indeterminate samples were positive by the second generation ELISAs. With few exceptions, all the RIBA II-negative samples were negative by the second generation ELISAs. Eleven blood donor sera were positive by test IV exclusively where RIBA II and other supplementary assays were negative. The recently introduced second generation anti-HCV ELISAs were found to have a higher sensitivity than the first generation test. The tests also showed a good concordance with the exception of test IV in the group of blood donor sera.
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PMID:A comparison between one first generation and three second generation anti-HCV ELISAs: an investigation in high- and low-risk subjects in correlation with recombinant immunoblot assay and polymerase chain reaction. 128 30

In May 1990 a specific enzyme immunoassay (EIA) for NANBH was developed by recombinant DNA technology which detects antibodies to a virus called hepatitis C virus (HCV). The anti-HCV EIA was manufactured by Ortho Diagnostic Systems with recombinant antigens from Chiron Corp. based on extraction from high infectious titer chimpanzee plasma RNA after transcription into cDNA. We tested the anti-HCV prevalence of blood donors and hemodialysis patients. The anti-HCV prevalence with the first generation test was 0.52% (Ortho), 0.87% (Abbott) in blood donors and 4.16% in hemodialysis patients. The second generation anti-HCV test (Abbott) with improved sensitivity and specificity comprises 0.25% repeated anti-HCV-positive blood donors and 8.2% anti-HCV-positive hemodialysis patients.
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PMID:[Decreasing the risk of post-transfusion non-A, non-B hepatitis by anti-HCV screening]. 128 60

Stored serum samples from 7,179 nonselected blood donors were tested for anti-HCV using Ortho EIA first generation. Results were compared to data acquired by anti-HBc testing and ALT levels found in routine testing. 24 donors (0.33%) were repeatedly reactive with Ortho HCV EIA, 230 (3.20%) were anti-HBc-positive and 138 (1.92%) had raised ALT levels > or = 36 IU/l. A low correlation was found between HCV antibody screening with EIA and surrogate testing. When tested in addition with the Abbott HCV EIA, 20 of the 24 Ortho EIA-positive subjects showed a positive reaction. In the Abbott neutralization test 13 of these 20 (65%) were reactive. 8 (33.33%) of the 24 Ortho-EIA-positive donors were positive in the two-antigen-RIBA (first generation), 8 were indeterminate and 8 were nonreactive. The neutralization test and the RIBA can be used as supplementary tests fo further analyze HCV-EIA-positive specimens.
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PMID:Detection of antibodies to hepatitis C virus in blood donors and their relationship to surrogate markers. 128 65

A study of a cross-section of the Hong Kong Chinese population was done to investigate the seroprevalence of hepatitis C. Healthy subjects were random visitors of a health exhibition, while clinical subjects were recruited from an outpatient liver disease clinic, sexually transmitted disease clinics, dialysis centres and drug rehabilitation centres. A total of 910 subjects were tested. The assay kits were from Abbott and Ortho laboratories. Of the general population, 0.5% was found to be positive for antibody to hepatitis C (anti-HCV). Suspected chronic non-A non-B patients, parenteral drug abusers and haemophiliacs shared a common high (up to 70%) prevalence of anti-HCV. Sexual partners of index patients, homosexuals and female prostitutes as well as hepatitis B carriers had 0% prevalence. It was concluded that parenteral and blood product exposures were the two main risk factors for hepatitis C.
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PMID:Prevalence of hepatitis C infection in Hong Kong. 131 65

The specificity of first-generation enzyme-linked immunosorbent assays (ELIAs) for antibody detection in individuals with hepatitis C virus (HCV) infection has been questioned in some pathological situations. We observed a surprisingly high prevalence of anti-HCV antibodies in alcoholic patients, and thus, false-positive reactions in anti-HCV tests were strongly suspected. The introduction of new epitopes, particularly a core protein, C22 (second-generation tests), seems to increase the sensitivity of anti-HCV detection. In order to study the specificity of the second-generation tests, 60 serum samples from alcoholic patients found to be positive by the first-generation anti-HCV ELISA (Ortho) were reexamined by a second-generation anti-HCV enzyme immunoassay (Abbott) and a recombinant immunoblot assay (RIBA II; Chiron). Fifteen serum samples gave contradictory results when they were tested by the two assays. We performed nested polymerase chain reactions (PCRs) to confirm that the discrepancies that we observed could be due to the presence of low levels of anti-HCV antibodies, which were detected by a more sensitive test, or to unspecific positive reactions. Nested PCR revealed the presence of HCV RNA sequences in all anti-HCV-positive sera or sera that were weakly positive by ELISA. Anti-HCV positive by RIBA II was always correlated with the presence of viral RNA in serum, but HCV RNA was detected in RIBA II-negative sera. These results indicate that the specificity of the second-generation tests is an important improvement but that an HCV infection can still persist without detectable antibodies. PCR remains the reference assay to clear up controversial serology results and to detect HCV infection in patients with no anti-HCV-detectable immune response.
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PMID:Detection of hepatitis C virus sequences in sera with controversial serology by nested polymerase chain reaction. 131 39

The reference range of serum alanine amino transferase (ALT) for the local population was established by testing 5,000 random voluntary Chinese blood donors of various age groups of both sexes. In addition, 1,769 serum samples with elevated ALT levels were also collected for anti-HCV assays using both the Abbott and Ortho anti-hepatitis C virus (HCV) assay kits. The relationship between serum ALT and anti-HCV tests was studied and the performances of both kits used were compared. It was found that while the prevalence of serum anti-HCV was 0.4% among hepatitis B surface antigen-negative donors with normal ALT, subjects with ALT between 2 and 3 standard deviations (SD) and greater than 3 SD above the mean level had respective prevalence of anti-HCV 3 and 9.5 times that of the normal ALT subjects. Both anti-HCV kits were found to identify in majority the same positive population among the different groups of subjects studied. In addition, it was observed that for subjects who were anti-HCV-positive, the higher the serum ALT level, the higher the mean anti-HCV ELISA ratio and this observation was similar for both anti-HCV kits used. We conclude that: (1) there is a direct relationship between serum ALT level and anti-HCV positivity by EIA; (2) there is a direct correlation between serum ALT level and anti-HCV ELISA ratio, and (3) both Abbott and Ortho anti-HCV kits perform similarly in the identification of positive serum samples.
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PMID:A study of hepatitis C virus antibodies and serum alanine amino transferase in blood donors in Hong Kong Chinese. 132 15

In a series of 385 sera obtained from volunteer blood donors positive for the first-generation hepatitis C virus assay (Ortho), the viral genome was detected by polymerase chain reaction (PCR) in 89 sera (23%). Most PCR-positive sera were found positive with the c100-3 neutralisation assay (Abbott) and by two second-generation enzyme immunoassays (Abbott, Ortho). However overall specificity of these assays was rather low. By immunoblotting (Innogenetics and Chiron/Ortho) the specificity could be considerably improved and the best correlation with carrier state was obtained when analysing the results for lane-specific reaction: all 89 viral carriers and only 9 other donors had antibodies against structural 'core' epitopes. From the present data we can conclude that in screening a volunteer blood donor population the confirmation of antibodies against 'core' epitopes by immunoblotting is strongly associated with viral carriage.
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PMID:Confirmation of hepatitis C virus positive blood donors by immunoblotting and polymerase chain reaction. 133 36

The aim of our study was to confirm by Recombinant Immunoblot Assay (RIBA) and by neutralization assay the repeat positive reactions found by two commercially available EIAs (Ortho and Abbott) when testing samples from volunteer blood donors, patients with chronic liver disease and with hepatocellular carcinoma. Our data show a high confirmatory rate among patients with chronic viral NANBH and HCC, while among donors and patients with CLD other than NANBH the percentage of presumptive EIA positive reactions confirmed by RIBA and/or neutralization assay is much lower. In our experience, the neutralization assay appears to be somewhat more sensitive than RIBA, especially when samples show low EIA optical densities.
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PMID:Confirmation of anti-HCV EIA reactivities by RIBA and neutralization assay among blood donors and patients with chronic liver disease and hepatocellular carcinoma. 133 25


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