Abstract:
BACKGROUND:Measurements of the ionic current flowing through nanometer-scale pores (nanopores) have been used to analyze single DNA and RNA molecules, with the ultimate goal of achieving ultrafast DNA sequencing. However, attempts at purely electronic measurements have not achieved the signal contrast required for single nucleotide differentiation. In this report we propose a novel method of optical detection of DNA sequence translocating through a nanopore. METHODS:Each base of the target DNA sequence is 1st mapped onto a 2-unit code, 2 10-bp nucleotide sequence, by biochemical conversion into Designed DNA Polymers. These 2-unit codes are then hybridized to complementary, fluorescently labeled, and self-quenching molecular beacons. As the molecular beacons are sequentially unzipped during translocation through a <2-nm-wide nanopore, their fluorescent tags are unquenched and are detected by a custom-built dual-color total internal reflection fluorescence (TIRF) microscope. The 2-color optical signal is then correlated to the target DNA sequence. RESULTS:A dual-color TIRFM microscope with single-molecule resolution was constructed, and controlled fabrication of 1-dimensional and 2-dimensional arrays of solid-state nanopores was performed. A nanofluidic cell assembly was constructed for TIRF-based optical detection of voltage-driven DNA translocation through a nanopore. CONCLUSIONS:We present a novel nanopore-based DNA sequencing technique that uses an optical readout of DNA translocating unzipping through a nanopore. Our technique offers better single nucleotide differentiation in sequence readout, as well as the possibility of large-scale parallelism using nanopore arrays.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Soni GV,Meller Adoi
10.1373/clinchem.2007.091231subject
Has Abstractpub_date
2007-11-01 00:00:00pages
1996-2001issue
11eissn
0009-9147issn
1530-8561pii
clinchem.2007.091231journal_volume
53pub_type
杂志文章abstract::We compared serum lipoprotein cholesterol concentrations in a randomly selected subsample (n = 406) population of children (ages 5 to 17 years), as measured by the heparin-Ca2+ and the heparin-Mn2+ precipitation methods. Statistical analysis showed significant differences between results by these methods, especially i...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-10-01 00:00:00
abstract:BACKGROUND:The single cell represents the basic unit of all organisms. Most investigations have been performed on large cell populations, but understanding cell dynamics and heterogeneity requires single-cell analysis. Current methods for single-cell analysis generally can detect only one class of analytes. METHODS:Re...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2012.191445
更新日期:2012-12-01 00:00:00
abstract::We have developed an automated latex agglutination test for Treponema pallidum (TPLA) for measurement of the concentration of treponemal antibodies in syphilitic sera. The assay needs neither a complicated pretreatment of the sera nor special techniques. Intra- and interassay precision studies showed high reproducibil...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-08-01 00:00:00
abstract:BACKGROUND:Vascular smooth muscle cells (SMCs) migrate from the arterial media to the intima in the progression of atherosclerosis, and dysfunction of SMCs leads to enhanced atherogenesis. A soluble form of the LDL receptor relative with 11 ligand-binding repeats (sLR11) is produced by the intimal SMCs, and the circula...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2009.127027
更新日期:2009-10-01 00:00:00
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::We analyzed pancreatic enzyme data from 508 patients with suspected pancreatitis by neural network analysis, by an Expert multirule generation protocol, and by receiver-operator characteristic (ROC) curve analysis of a single test result. Neural network analysis showed that use of lipase provided the best means for di...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-09-01 00:00:00
abstract::We immunoenzymometrically measured creatine kinase (CK) isoenzyme MB in extracts of myocardium and in homogenates of five different skeletal muscles. CK-MB concentrations in the former averaged 80.9 micrograms/g wet tissue; in the skeletal muscles it varied widely, being (e.g.) 25-fold greater in diaphragm than in pso...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-02-01 00:00:00
abstract::The measurement of glycohemoglobin (GHb) by boronate affinity chromatography is useful in monitoring long-term glucose control in diabetic subjects. The inherent disadvantage of this method is the hands-on time required because the hemoglobin fractions are separated on individual disposable columns. To overcome this d...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-01-01 00:00:00
abstract:BACKGROUND:Cardiac troponins are specific biochemical markers of myocardial injury used in the diagnosis of acute myocardial disease and cardiac risk stratification. To avoid misclassification of patients, troponin assays must demonstrate precision at the low end of the measuring range. We report our evaluation of the ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2005.057216
更新日期:2006-02-01 00:00:00
abstract::The commonly used methods of assessing the precision of the automated leukocyte differential have certain drawbacks that affect the validity and comparability of results. In the present report, we introduce a procedure based on building precision profiles from a large number of within-run imprecision experiments. The ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-07-01 00:00:00
abstract::We describe use of near-infrared reflectance spectroscopy (NIRRS) to estimate the amount of fat in feces, for diagnosis of steatorrhea. After sample homogenization, the spectrum of the fecal homogenate is scanned over the near-infrared region. Assay of 94 samples of feces having a known concentration of fat showed the...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-04-01 00:00:00
abstract::Histograms of the results of quantitative clinical laboratory analyses are usually broadly skewed because some of the patients have normal values with respect to the analyte being measured while values for others are below or above normal. We describe a technique for calculating the maximum possible number of individu...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-04-01 00:00:00
abstract::We measured the concentration of carboxyhemoglobin (HbCO) in blood samples from 32 neonates by spectrophotometry (IL282 CO-Oximeter) and gas chromatography, finding a strong positive correlation (r = 0.89) between the concentration of fetal hemoglobin (Hb F) and HbCO as measured by spectrophotometry, but not by gas ch...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-05-01 00:00:00
abstract::The Center for Disease Control (CDC), the New York State Department of Health (NYSDH), the College of American Pathologists, and 23 manufacturers of diagnostic products participated in an interlaboratory study of aspartate aminotransferase (EC 2.6.1.1) methodologies. Six different lyophilized materials were prepared a...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-06-01 00:00:00
abstract::The new oral formulation of cyclosporine (CsA), Neoral (CsA-N), results in increased area under the curve (AUC) and decreased intra- and interindividual variation in blood concentrations and other pharmacokinetic (PK) parameters when compared with the current Sand-immune (CsA-S) formulation. The present study examines...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1997-01-01 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::This solid-phase competitive europium immunoassay of progesterone in plasma relies on antigen labeling. With this new approach, time-resolved fluoroimmunoassay can attain sensitivity and precision similar to that of conventional radioimmunoassay. The europium-labeling involves coupling diethylenetriaminepentaacetic ac...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-03-01 00:00:00
abstract:BACKGROUND:Oxidation of lipids in lipoproteins and cells may initiate and enhance the early development of cardiovascular disease. METHOD AND RESULTS:We assayed F(2)-isoprostanes, oxidation products of arachidonic acid, by gas chromatography-mass spectrometry in a biracial cohort of 2850 young healthy adult men and wo...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2004.037630
更新日期:2005-01-01 00:00:00
abstract::We evaluated a new discrete analyzer, the Hitachi 705. The instrument can process sequentially as many as 19 different enzymes or other analytes, with a throughput of 180 tests per hour. We evaluated the spectrophotometer, the sample and reagent probes, and the performance for 11 assays: creatine kinase, aspartate ami...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-04-01 00:00:00
abstract::Based on the quantitative determination of creatinine, total protein, albumin, alpha 1-microglobulin, IgG, alpha 2-macroglobulin, and N-acetyl-beta, D-glucosaminidase in urine in combination with a test strip screening, the findings of hematuria, leukocyturia, and proteinuria can be assigned to prerenal, renal, or pos...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-08-01 00:00:00
abstract::This procedure for routine quantification of albumin in urine is based on the dye-binding properties of albumin with bromphenol blue. The absorbance of 100 microL of urine mixed with 3 mL of color reagent is measured against blank reagent at 610 nm after 30 s. Results vary linearly with albumin concentration up to 6 g...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-02-01 00:00:00
abstract::We evaluated the Reflotron (Boehringer Mannheim) system for measuring activities of alanine aminotransferase (EC 2.6.1.2) and gamma-glutamyltransferase (EC 2.3.2.2). Within- and between-day precision were comparable with conventional analytical methods. The linear range of the methods extended to more than 40-fold the...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-06-01 00:00:00
abstract::We describe an enzymic assay for the functional activity of alpha 1-antitrypsin, in which tosyl-Gly-Pro-Lys-p-nitroanilide acetate (Chromozym PL) is the trypsin substrate. For the indicating reaction, the concentration of substrate is about double the Km of its reaction with trypsin. This assay is an improvement over ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-11-01 00:00:00
abstract::A new computer-interfaced system of general applicability in the clinical laboratory has been built. The system is constructed around a new type multicuvette. Quantitation is currently by spectrophotometry and is on-line, with use of a mini-computer with output going to an electrostatic printer/plotter. The system can...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-08-01 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::Intra- and interindividual components of biological variation have been determined for total thyroxin (TT4), free thyroxin (FT4), total triiodothyronine (TT3), free triiodothyronine (FT3), and thyrotropin (TSH). Calculated analytical goals (CV, %) for the precision required for optimal patient care are: TT4 less than ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-06-01 00:00:00
abstract::Bush and Reed reported (Clin Chem 1987;33:821-3) that the reaction of albumin with bromcresol purple but not with bromcresol green underestimated the concentration of albumin in synthetically obtained bilirubin-albumin (Bd) by 29%. Their unproven assumption was that chemically synthesized Bd behaved in a manner indist...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-07-01 00:00:00
abstract:BACKGROUND:Extracellular matrix alterations are important elements in the arterial changes seen in diabetes, being associated with increased vascular stiffness and the development of cardiovascular diseases. However, no biomarkers for diabetes-related arterial changes have been defined. METHODS:Mammary artery specimen...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2011.162966
更新日期:2011-11-01 00:00:00
abstract::We developed a solid-phase two-site immunoenzymometric assay (IEMA) of the estrogen-induced 52-kDa cathepsin D (EC 3.4.23.5) and its processed forms (48-kDa and 34-kDa proteins) in cytosols of breast cancer tissues, using two monoclonal antibodies directed against two different epitopes of these antigens. The first an...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-01-01 00:00:00
abstract:BACKGROUND:Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with vari...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2016.259036
更新日期:2017-04-01 00:00:00