Abstract：
BACKGROUND:In malignancy, gene mutations frequently occur in tumor suppressor genes such as p53 and are sporadically located. We describe a homogeneous method for amplification and mutation scanning, and apply the method to the p53 gene. METHODS:Using a series of overlapping fluorescein-labeled oligonucleotides complementary to a wild-type p53 sequence, we detected somatic mutations in colorectal cancers by aberrant probe:target melting temperatures (T(m)). The probes were designed so that fluorescence decreased on target annealing as a result of deoxyguanosine quenching. Probes were walked along the sequence to be scanned, using two to three probes per cuvette and placing overlapping probes in separate reactions. After amplification, the reaction was cooled to anneal probes and then slowly heated (0.1 degrees C/s) while fluorescence was continuously monitored. Somatic mutations in tumor tissue were detected by changes from a characteristic wild-type melting curve profile using leukocyte DNA. RESULTS:A complete scanning of the DNA binding domain (exons 5-8) of the p53 gene was completed in a single run ( approximately 30 min) starting from genomic leukocyte DNA. To show proof-of-principle, p53 exons 6-8 from 63 colon cancers were probe-scanned and showed 100% agreement with direct sequencing for detecting alterations from wild-type DNA. CONCLUSIONS:p53 mutation scanning by single-labeled hybridization probes is a homogeneous, rapid, and sensitive method with application in both research and clinical diagnostics.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Millward H,Samowitz W,Wittwer CT,Bernard PSsubject
Has Abstractpub_date
2002-08-01 00:00:00pages
1321-8issue
8eissn
0009-9147issn
1530-8561journal_volume
48pub_type
杂志文章abstract：:Time-resolved fluoroimmunoassay was developed for the detection of synovial-type phospholipase A2 (s-PLA2) in human serum. This solid-phase, sandwich assay uses a polyclonal rabbit antibody raised against synovial-type group II PLA2 produced in Escherichia coli. No cross-reactions were detected between s-PLA2 and PLA2...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-09-01 00:00:00
abstract：:This highly sensitive method for determining retinol-binding protein in human serum and urine is based on a double-antibody "sandwich"-type enzyme-linked immunosorbent assay. The assayable concentration range is 0.8-48 micrograms/L, the detection limit 0.2 micrograms/L. Within-assay coefficients of variation for 10 de...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-01-01 00:00:00
abstract：:We evaluated the EMIT Cyclosporine Assay (Syva Co., Palo Alto, CA), using the Cobas-Mira analyzer to assess the precision, accuracy, and analytical recovery from whole-blood samples supplemented with cyclosporine. We also performed comparative analysis of whole-blood samples containing cyclosporine from liver and kidn...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-12-01 00:00:00
abstract：:Various laboratories have reported endogenous digoxin-like immunoreactive factor(s) (DLIF) in blood from patients in renal failure or liver failure, from newborn infants, and from third-trimester pregnant women. Similar immunoreactivity has been detected in amniotic fluid, in cord blood, and in urine and serum from no...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1985-09-01 00:00:00
abstract：:We have developed a new assay for cortisone (E) in serum, saliva, and urine involving Celite chromatography followed by RIA with 125I-labeled E and scintillation proximity assay. The chromatography step separates cortisol (F) from E, and in combination with their RIAs, permits assessment of the status of the F-E shutt...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-08-01 00:00:00
abstract：:Using results from patients, we studied several methods for long-term (years) monitoring of the analytical stability of various chemistry tests. For each test, we obtained on a bimonthly basis histograms of > or = 200 to approximately 2000 data points and determined the median, mean, and mean of those data points in t...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1996-06-01 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：BACKGROUND:Paraoxonase may protect lipoproteins and cell membranes from peroxidation, and alterations in the activity of this enzyme have been associated with some chronic diseases. Serum paraoxonase appears to be mainly under genetic control, but some studies suggest that environmental factors may also modulate its ac...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.9.1491
更新日期：2003-09-01 00:00:00
abstract：:We describe optimal conditions for immobilization of two monoclonal antibodies to progesterone for solid-phase assays. Polystyrene surfaces are refined with Protein A to achieve uniform, reproducible, stable, and sterically accessible immobilization of immunoglobulins (IgG). To this end, we optimized the amount of imm...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-08-01 00:00:00
abstract：BACKGROUND:For the past 30 years, the Sanger method has been the dominant approach and gold standard for DNA sequencing. The commercial launch of the first massively parallel pyrosequencing platform in 2005 ushered in the new era of high-throughput genomic analysis now referred to as next-generation sequencing (NGS). ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2008.112789
更新日期：2009-04-01 00:00:00
abstract：:A high (2.0 mol/L) NaCl concentration apparently suppresses the secondary-site binding of bilirubin by human serum albumin. Thus if an excess of bilirubin is added to human serum albumin or to neonatal serum in buffer containing 0.1 mol of tris(hydroxymethyl)aminomethane and 2.0 mol of NaCl per liter (pH 7.5), and the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-01-01 00:00:00
abstract：:We describe a liquid-chromatographic assay for AZT in human plasma and urine. This assay involves the use of two internal standards, allowing reference of AZT peaks to the appropriate internal standard, the choice depending on the range of concentrations encountered. This method is isocratic, specific, sensitive enoug...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-08-01 00:00:00
abstract：:We have used a monoclonal antibody-based ELISA for plasma prothrombin fragment 1.2 (F1.2) to establish appropriate sample collection and storage conditions for this biomarker of thrombin generation. F1.2 concentrations were not altered by exogenous factor Xa, thrombin, or thromboplastin if blood was collected by routi...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-10-01 00:00:00
abstract：:Endotoxin, the lipopolysaccharide cell wall constituent of Gram-negative bacteria, produces symptoms of the Gram-negative sepsis syndrome. By measuring endotoxin in blood from septic patients it may be possible to select a subpopulation of patients in which mortality can be prevented by treatment with anti-endotoxin a...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-08-01 00:00:00
abstract：:Hepatitis A virus (HAV) infection occurs worldwide and is an important cause of acute viral hepatitis in the US. In this review, I cover the epidemiology, course of infection, clinical manifestations, serological responses, and prevention of this infection. Although most patients completely recover from this disease, ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1997-08-01 00:00:00
abstract：:The isoenzyme pattern of N-acetyl-beta-glucosaminidase (NAG) in serum and urine was studied in two groups of patients with diabetes mellitus and in 30 control subjects. Total NAG activity was significantly (P less than 0.001) increased in the serum and urine of the 20 diabetics with vascular complications, but was ins...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-12-01 00:00:00
abstract：:We evaluated the performance of two homogeneous assays for quantifying HDL cholesterol (HDL-C) and compared them with the phosphotungstic acid (PTA)/ MgCl2 assay. Both homogeneous HDL-C assays were precise, having a within-run CV of < 1.20% and a between-run CV of < 4.07%. The HDL-C values (y) measured by the two homo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-06-01 00:00:00
abstract：:The aim of this investigation was to study the distribution of arsenic species in human organs following fatal acute intoxication by arsenic trioxide. The collected autopsy samples of most organs were ground and dried, and the total arsenic was measured by electrothermal atomic absorption spectrometry (ETAAS). The ars...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1999-02-01 00:00:00
abstract：BACKGROUND:Determinations of certain forms of prostate-specific antigen (PSA) have been shown to increase the specificity for prostate cancer (PCa). One such variant, proteolytically active PSA, is a potentially useful tumor marker, but it is not specifically recognized by antibodies. Using phage display libraries, we ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2003.026146
更新日期：2004-01-01 00:00:00
abstract：:Market forces have dramatically influenced the environment in which healthcare is delivered, but these changes do not need to be interpreted negatively by community laboratorians. Only total vertical integration of laboratory medicine can control episode-of-care cost. Opportunities also exist for horizontal integratio...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1996-05-01 00:00:00
abstract：:Using a continuous-flow (Technicon SMAC) analyzer, we measured the concentration of 20 analytes in capillary blood sera from 149 neonates in an intensive-care unit. Included were term and pre-term babies, both groups with and without asphyxia. The values obtained, especially for those enzymes and analytes that are rel...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-04-01 00:00:00
abstract：BACKGROUND:The susceptibility of plasma lipids to oxidation is thought to be a factor contributing to atherogenic risk. Various groups have studied the in vitro oxidizability of isolated LDL and examined the effects of conventional antioxidants. The drawbacks associated with the isolation of LDL for evaluation of in vi...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.051078
更新日期：2005-11-01 00:00:00
abstract：BACKGROUND:5α-Reductase 2 deficiency (5ARD) is a known cause of 46,XY disorders of sex development (DSD). Traditionally, the diagnosis relies on dihydrotestosterone (DHT) measurement, but the results are often equivocal, potentially leading to misdiagnosis. We reviewed alternative approaches for diagnosis of 5ARD. MET...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2012.196501
更新日期：2013-05-01 00:00:00
abstract：:We compared the performance of three methods for quantifying high-density lipoprotein cholesterol (HDL-C) with the Reference Method for HDL-C, using samples with a wide range of triglyceride (TG) concentrations (290-18000 mg/L). All three comparison assays-- utilizing a magnetic dextran sulfate precipitating reagent, ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1996-05-01 00:00:00
abstract：:Investigating the interference of danazol in testosterone radioimmunoassays, we measured serum testosterone concentrations in the serum of women receiving danazol, with three testosterone radioimmunoassay kits. All of these showed positive interference by danazol or its metabolites (or both). Values for testosterone o...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-01-01 00:00:00
abstract：:We describe a rapid polarization fluoroimmunoassay for screening for the presence of barbiturates in urine. The single reagent is prepared by pre-mixing antiserum with a fluorescein-labeled barbiturate derivative as tracer. Use of a mixture of two sheep antisera, raised against different barbiturate immunogens, result...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-11-01 00:00:00
abstract：BACKGROUND:Absolute plasma hepcidin concentrations measured by various procedures differ substantially, complicating interpretation of results and rendering reference intervals method dependent. We investigated the degree of equivalence achievable by harmonization and the identification of a commutable secondary refere...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2016.256768
更新日期：2016-07-01 00:00:00
abstract：:We have developed a series of photochemical silver stains that are more than 100 times as sensitive as the Coomassie Blue stains. These stains facilitate the quantitative estimation of proteins in two-dimensional gel patterns. Recently, we discovered that by using light (photodevelopment) to catalyze the reduction of ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-12-01 00:00:00
abstract：:To see if saliva is a valid substitute for plasma in assay of progesterone even when concentrations of hormone and binding proteins are fluctuating, we determined the concentrations of total and free progesterone in plasma and salivary progesterone in specimens from 36 women volunteers during the course of pregnancy a...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-01-01 00:00:00
abstract：:We examined the mechanism by which hypertriglyceridemia interferes with the Roche Diagnostics "Amylochrome" procedure for measurement of amylase in lactescent samples with normal and above-normal amylase activity. The serum blank recommended in the Amylochrome protocol to compensate for lactescence remains inappropria...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-10-01 00:00:00
