Abstract：
BACKGROUND:Rapid detection of biological threat agents is critical for timely therapeutic administration. Fluorogenic PCR provides a rapid, sensitive, and specific tool for molecular identification of these agents. We compared the performance of assays for 7 biological threat agents on the Idaho Technology, Inc. R.A.P.I.D., the Roche LightCycler, and the Cepheid Smart Cycler. METHODS:Real-time PCR primers and dual-labeled fluorogenic probes were designed to detect Bacillus anthracis, Brucella species, Clostridium botulinum, Coxiella burnetii, Francisella tularensis, Staphylococcus aureus, and Yersinia pestis. DNA amplification assays were optimized by use of Idaho Technology buffers and deoxynucleotide triphosphates supplemented with Invitrogen Platinum Taq DNA polymerase, and were subsequently tested for sensitivity and specificity on the R.A.P.I.D., the LightCycler, and the Smart Cycler. RESULTS:Limit of detection experiments indicated that assay performance was comparable among the platforms tested. Exclusivity and inclusivity testing with a general bacterial nucleic acid cross-reactivity panel containing 60 DNAs and agent-specific panels containing nearest neighbors for the organisms of interest indicated that all assays were specific for their intended targets. CONCLUSION:With minor supplementation, such as the addition of Smart Cycler Additive Reagent to the Idaho Technology buffers, assays for DNA templates from biological threat agents demonstrated similar performance, sensitivity, and specificity on all 3 platforms.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Christensen DR,Hartman LJ,Loveless BM,Frye MS,Shipley MA,Bridge DL,Richards MJ,Kaplan RS,Garrison J,Baldwin CD,Kulesh DA,Norwood DAdoi
10.1373/clinchem.2005.052522subject
Has Abstractpub_date
2006-01-01 00:00:00pages
141-5issue
1eissn
0009-9147issn
1530-8561pii
52/1/141journal_volume
52pub_type
杂志文章abstract：:We describe a time-resolved immunofluorometric assay (IFMA) for corticotropin in unextracted human plasma, based on the use of two monoclonal antibodies: europium-labeled antibody 1A12 and antibody 2A3 coated onto microtiter wells. We compared the results of this assay with those of an immunoradiometric assay (IRMA) p...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-10-01 00:00:00
abstract：:A precolumn derivatization method was optimized for rapid and specific analysis of total urinary hydroxyproline by HPLC. After an overnight hydrolysis, urine samples dried and reconstituted with the internal standard cysteic acid (in sodium hydrogen carbonate, pH 9.3) were derivatized with N,N-diethyl-2,4-dinitro-5-fl...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-03-01 00:00:00
abstract：:Two-hour postprandial specimens have a -14% proportional bias for lipoprotein(a) [Lp(a)], a -0.035 g/L systematic bias for apolipoprotein (apo) A-I, and a -9% proportional bias for apo B, compared with values in 12-h fasting specimens. Although a physiological hemodilution appears to account for a portion of these bia...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-03-01 00:00:00
abstract：:alpha 1-Antitrypsin (A1AT) deficiency is a relatively common autosomal recessive disease, resulting most often from a single base pair (1 bp) substitution called the Z mutation. Previous genetic tests for carriers and affected patients have relied on quantitative binding of radioactive probes to an amplified gene prod...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-08-01 00:00:00
abstract：:The objective of this study was to develop a clinical laboratory method for subclass typing of human immunoglobulin G (IgG) paraproteins. Serum proteins were isoelectrically focused (IEF) in a mini-gel and passively blotted by capillary diffusion onto untreated nitrocellulose. Unreacted sites on the nitrocellulose wer...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-03-01 00:00:00
abstract：:A modified heparin-Sepharose affinity chromatography procedure (Boberg et al., J. Lipid Res. 18:544-547, 1977) was developed to determine two different triglyceride lipase activities in human post-heparin plasma: hepatic triglyceride lipase (I) and lipoprotein lipase (II). With this procedure, lipoproteins were separa...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-05-01 00:00:00
abstract：:A two-site assay was developed by use of the "dissociation and enhancement lanthanide fluoroimmunoassay" (DELFIA) technique for determination of salmon calcitonin (SCT) in serum after administration to osteoporotic patients. Polyclonal antibodies were produced in rabbits immunized with SCT coupled to ovalbumin. After ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-09-01 00:00:00
abstract：:For 51 liver allograft recipients, we evaluated whether serum profiles of delta (Bd) and conjugated bilirubins (Bc) could be used to diagnose rejection during the first 30-50 postoperative days, in comparison with histology as the "gold standard." Daily measurements of aspartate aminotransferase, alkaline phosphatase,...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1990-01-01 00:00:00
abstract：:We describe a system for detection of leukemia cells involving complement-mediated cytotoxic reaction and an image processing system, consisting of a charge-coupled-device image sensor, an image memory board, a personal computer, and a phase-contrast microscope. Then added to a cell suspension, monoclonal antibody spe...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-04-01 00:00:00
abstract：:This improved assay of rheumatoid factors in serum, described here for use with the Baker "Encore" centrifugal analyzer, is efficient, with 250-sample throughput per hour; reproducible, with between-batch CV = 5% and within-batch CV = 2% (mid-assay range); and results correlate well (r = 0.9) with those by other metho...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-02-01 00:00:00
abstract：BACKGROUND:Specific gravity (SG) may perform as well as creatinine (CR) correction for adjusting urinary hormone concentrations, as well as offer some advantages. We compared the two methods and applied them to US and Bangladeshi specimens to evaluate their use in different populations. METHODS:Pearson correlations be...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2004.032292
更新日期：2004-05-01 00:00:00
abstract：:We studied the effects of some chelators on creatine kinase activity. Creatine kinase is competitively inhibited by endogenous polyvalent cations (e.g., calcium, Ki = 4.5 mmol/L); this can be reversed by adding chelators to the reagent, resulting in a mean increase in activity of 1.14-fold at 25 degrees C and 1.18-fol...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-03-01 00:00:00
abstract：:Taurine was measured in cerebrospinal fluid by reacting it with fluorescamine to form a fluorescent derivative, followed by separation on a reversed-phase column and fluorometric detection and evaluation. The assay is rapid (17 min) and sensitive to as little as 1 mumol/L. The mean value for 27 cerebrospinal fluid sam...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-08-01 00:00:00
abstract：BACKGROUND:Little research has been done to examine whether gamma-glutamyltransferase (GGT) is prospectively associated with the development of chronic kidney disease (CKD). We performed a prospective study to examine the association between GGT and the risk for the development of CKD. METHODS:The study cohort include...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2006.078980
更新日期：2007-01-01 00:00:00
abstract：BACKGROUND:Kallikrein-related peptidase 6 (KLK6), a member of the serine protease family of kallikrein (KLK) genes, is dysregulated in ovarian carcinomas (OCa) and its overexpression is associated with poor prognosis. Regulation of its expression is poorly understood and is likely to be influenced by multiple mechanism...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.193060
更新日期：2013-01-01 00:00:00
abstract：:We evaluated the analytical performance of new estradiol and progesterone assays performed on the Bayer Immuno 1 system. Within-run and between-day CVs for estradiol at concentrations of 116.8-6645.8 pmol/L were < or = 6.4% and for 5.54-103.95 nmol/L progesterone were < or = 7.7%, thus meeting published analytical goa...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-09-01 00:00:00
abstract：:A fixed-time incubation method for measuring acetylsalicylic acid esterase was improved by using a higher concentration of the substrate, acetylsalicylic acid, and an activator, Ca2+. The enzymatic activity in serum was 25-fold that measured by earlier methods. Inhibition studies showed a pattern similar to that repor...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-03-01 00:00:00
abstract：:The Axial Separation Module (ASM), which separates whole-blood specimens serially in Axial Process Containers (APC), was evaluated for clinical performance at the University of Virginia Health Sciences Center (UVA HSC) in a community-based outpatient laboratory (North Ridge Clinic). We hypothesized that moving the tas...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-02-01 00:00:00
abstract：:Patients' sera were analyzed for carbamazepine and ethosuximide by enzyme immunoassay (x) and gas chromatography (y), and the results were compared. The correlation coefficients were: for carbamazepine, x vs. y 0.94 (n = 93); for ethosuximide, x vs. y 0.99 (n = 30). These results suggest that the two methods could be ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-09-01 00:00:00
abstract：BACKGROUND:Blood concentrations of the calcineurin inhibitors (CNIs) cyclosporine and tacrolimus are currently measured to monitor immunosuppression in transplant patients. The measurement of calcineurin (CN) phosphatase activity has been proposed as a complementary pharmacodynamic approach. However, determining CN act...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2013.213264
更新日期：2014-02-01 00:00:00
abstract：:Establishing associations between environmental agents and disease presents challenges to both epidemiologists and toxicologists, particularly in cases of complex gene-environment interactions and when there is a long latency between exposure and disease. Biologic markers, physiological signals that reflect exposure, ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1994-07-01 00:00:00
abstract：:We have developed a fully enzymatic method to measure 1,5 anhydro-D-glucitol (1,5-AG) in serum through use of pyranose oxidase (PROD: EC 1.1.3.10), glucokinase (EC 2.7.1.2), and an ATP-regenerating system. In a previous report (Clin Chem 1989;35:2039-43) the glucose interfering with the measurement of 1,5-AG was remov...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-11-01 00:00:00
abstract：:A colorimetric method for measuring anti-NAD+ glycohydrolase in human sera has been developed. The assay involves the inhibition of NAD+ glycohydrolase (EC 3.2.2.5) by the antibody and determination of the noninhibited enzyme activity by using an enzymatic amplifying system for NAD+. The assay is easily carried out an...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-06-01 00:00:00
abstract：:We studied several risk factors in relation to parental cardiovascular heart disease: total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, apolipoprotein (apo) A-I, apo B, and lipoprotein(a) [Lp(a)] were determined in 322 serum samples (43 from subjec...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-03-01 00:00:00
abstract：:We assessed the LKB "Delfia" (time-resolved dissociation-enhanced lanthanide fluoroimmunoassay) and the Amersham "Amerlite" (enhanced luminescent immunometry) assays of thyrotropin in serum. Both assays are sensitive (respective detection limits: 0.02 and 0.04 milli-int. unit/L) and have very good within- and between-...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-08-01 00:00:00
abstract：:We describe microplate methods for measurement of human urinary albumin (HUA) by competitive enzyme-linked immunosorbant assay (ELISA) and creatinine with a modified commercial enzymatic kit. Incorporation of substrate mixing into the competitive ELISA changes the dynamic absorbance-concentration response, greatly sim...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-07-01 00:00:00
abstract：:Since 1991, the Ontario Laboratory Proficiency Testing Program has assessed the analytical performance of creatine kinase (CK; EC 2.7.3.2) isoenzyme-2, using fresh human serum supplemented with purified human CK isoenzymes. In Ontario, the 142 laboratories licensed to analyze CK-2 use a variety of methods: electrophor...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-02-01 00:00:00
abstract：:We describe an enzymatic fluorometric method for determining glucose concentrations in blood samples by analysis on a semi-solid surface (silicone-rubber pads). The method is an adaptation of existing spectrophotometric procedures based on the sequential coupling reactions with glucose-6-phosphate dehydrogenase and NA...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1975-11-01 00:00:00
abstract：:Circulating prostate cells can be detected in cancer patients by using reverse transcriptase-PCR (RT-PCR) assay for prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSM) mRNA. A quality-control study involving a conventional RT-PCR assay was performed and, surprisingly, detected both transcripts...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1998-03-01 00:00:00
abstract：:We describe a mechanized chromogenic assay for factor X, the results of which correlate well with those for the one-stage clotting assays for factor X in which it is activated either via the extrinsic pathway by thromboplastin or directly by Russell's viper venom. We purified human factor X and raised monospecific ant...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-06-01 00:00:00
