Abstract：
BACKGROUND:First introduced in 2006, recombinase polymerase amplification (RPA) has stirred great interest, as evidenced by 75 publications as of October 2015, with 56 of them just in the last 2 years. The widespread adoption of this isothermal molecular tool in many diagnostic fields represents an affordable (approximately 4.3 USD per test), simple (few and easy hands-on steps), fast (results within 5-20 min), and sensitive (single target copy number detected) method for the identification of pathogens and the detection of single nucleotide polymorphisms in human cancers and genetically modified organisms. CONTENT:This review summarizes the current knowledge on RPA. The molecular diagnostics of various RNA/DNA pathogens is discussed while highlighting recent applications in clinical settings with focus on point-of-care (POC) bioassays and on automated fluidic platforms. The strengths and limitations of this isothermal method are also addressed. SUMMARY:RPA is becoming a molecular tool of choice for the rapid, specific, and cost-effective identification of pathogens. Owing to minimal sample-preparation requirements, low operation temperature (25-42 °C), and commercial availability of freeze-dried reagents, this method has been applied outside laboratory settings, in remote areas, and interestingly, onboard automated sample-to-answer microfluidic devices. RPA is undoubtedly a promising isothermal molecular technique for clinical microbiology laboratories and emergence response in clinical settings.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Daher RK,Stewart G,Boissinot M,Bergeron MGdoi
10.1373/clinchem.2015.245829subject
Has Abstractpub_date
2016-07-01 00:00:00pages
947-58issue
7eissn
0009-9147issn
1530-8561pii
clinchem.2015.245829journal_volume
62pub_type
杂志文章,评审abstract：:We describe an enzyme-linked immunosorbent assay (ELISA) for urinary albumin. It requires only commercially available reagents, can detect as little as 16 micrograms of albumin per liter, and analytical recovery ranges from 92 to 116%. The assay is simple, rapid, and inexpensive. Albumin excretion was 6.2 (SD 4.1) mg/...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-04-01 00:00:00
abstract：:We present an improved fructosamine assay for use with the Technicon RA-1000 analyzer. Features of this assay include: specimen throughput of 100 per hour, within-batch CV of 1.5%, between-batch CV of 2.4%, standard curve linear up to 24.9 mmol of fructosamine per liter, and good correlation (r = 0.82) with hemoglobin...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-12-01 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：BACKGROUND:In vitro and animal studies suggest that tumor necrosis factor alpha (TNF-alpha) modulates intestinal iron transport. We hypothesized that the effect of TNF-alpha might be particularly relevant if iron absorption is not effectively controlled by the HFE gene. METHODS:In patients with homozygous C282Y hemoch...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.065417
更新日期：2006-08-01 00:00:00
abstract：BACKGROUND:We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets. METHODS:In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector ol...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1999-09-01 00:00:00
abstract：:We describe the measurement of rheumatoid factor in human sera with a rate nephelometer. The National Reference Preparation for Rheumatoid Factors is used to calibrate the assay in International Units. We used Hyland Positive Control, Level I, as a secondary standard. The standard curve is exponential, but is linear w...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-11-01 00:00:00
abstract：:We describe a solid-phase radioimmunometric sandwich assay for a new tumor marker defined by a monoclonal antibody (19-9). This antibody reacts with a carbohydrate antigenic determinant (CA 19-9) found at low concentrations in sera from healthy individuals but frequently increased in sera from patients with adenocarci...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-03-01 00:00:00
abstract：:Immunoradiometric analysis has made it possible to measure ferritin reliably in serum. The determination is a sensitive means of assessing body iron because concentrations of ferritin closely parallel the body's reticuloendothelial iron stores. The information provided is similar to that obtained from bone-marrow aspi...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-01-01 00:00:00
abstract：BACKGROUND:A unique requirement for the molecular diagnosis of mitochondrial DNA (mtDNA) disorders is the ability to detect heteroplasmic mtDNA mutations and to distinguish them from homoplasmic sequence variations before further testing (e.g., sequencing) is performed. We evaluated the potential utility of temporal te...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章
doi：
更新日期：1999-08-01 00:00:00
abstract：:To assess the distribution of serum concentrations of the atherogenic lipoprotein(a) [Lp(a)] in an obese population and the possible effects of weight reduction, we determined Lp(a) in 52 white, moderately obese subjects (16 men and 36 women). The subjects were participating in a weight-reduction program of diet, exer...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章
doi：
更新日期：1991-07-01 00:00:00
abstract：:We measured immunoglobulins in the sera of 33 patients on days 1, 3, 6, 10, and 17 and three to four weeks after surgical operations (mostly hysterectomy or appendectomy) or (six patients) after spinal injury. In the absence of infection or blood transfusion, IgG usually decreased slightly and transiently after hyster...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1975-05-01 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：BACKGROUND:Plant-derived cardenolides reportedly possess anticancer properties in human leukemic cells via selective induction of apoptosis, cell cycle arrest, and differentiation. Selective induction of apoptosis with mammalian-derived digoxin-like immunoreactive factor (DLIF) could provide new strategies for anticanc...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2006.082081
更新日期：2007-07-01 00:00:00
abstract：:Throughout the 1970s, controversy centered both on immunoassay "sensitivity" per se and on the relative sensitivities of labeled antibody (Ab) and labeled analyte methods. Our theoretical studies revealed that RIA sensitivities could be surpassed only by the use of very high-specificity nonisotopic labels in "noncompe...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1991-11-01 00:00:00
abstract：:Zinc protoporphyrin is the predominent fluorescent porphyrin accumulating in erythrocytes as a result of chronic lead absorption or iron-deficiency anemia. Although diagnostic concentrations or thresholds for it in erythrocytes have been inferred, normal adult averages have not been established. We quantitatively asse...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-09-01 00:00:00
abstract：:A simple, direct radioimmunoassay for cortisol in human serum and plasma is described. An antiserum, raised in sheep to a cortisol-3-(O-carboxymethyl)oxime/bovine serum albumin conjugate, is coupled to microcellulose. No extraction is required because plasma samples and standards are incubated with the antiserum and a...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-05-01 00:00:00
abstract：:We have studied factors affecting biological variation in total plasma alkaline phosphatase in a population of 32 329 apparently healthy subjects four years old or older. Quantification of the bone and liver isoenzymes after thermal denaturation made it possible to specify the contributions of each isoenzyme to variat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-04-01 00:00:00
abstract：:We detected the presence of creatine kinase (EC 2.7.3.2) MB isoenzyme (CK-MB) in serum by using an immunoinhibition technique to inactivate the creatine kinase isoenzyme of muscle origin (CK-MM). The GEMSAEC centrifugal analyzer is ideally suited for this procedure because it rapidly mixes reagents and has a throughpu...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-04-01 00:00:00
abstract：BACKGROUND:The determination of cellular beta-galactocerebrosidase activity is an established procedure to diagnose Krabbe disease and monitor the efficacy of gene/stem cell-based therapeutic approaches aimed at restoring defective enzymatic activity in patients or disease models. Current biochemical assays for beta-ga...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.115873
更新日期：2009-03-01 00:00:00
abstract：:To validate azotemic markers as an index for intradialytic changes in solute concentration, we compared eight solutes (pseudouridine, xanthine, hypoxanthine, peak 4, peak 5, p-hydroxyhippuric acid, indoxyl sulfate, and hippuric acid) with five classical azotemic markers (urea, creatinine, uric acid, phosphate, and pot...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-08-01 00:00:00
abstract：BACKGROUND:The discovery of cell-free fetal DNA in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis and monitoring. Among the fetal markers that have been described, methylation markers are sex and polymorphism independent. Methylation-sensitive restriction endonucleases are commonly u...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2007.092619
更新日期：2007-11-01 00:00:00
abstract：BACKGROUND:We evaluated assays to measure both total tissue plasminogen activator (tPA) and the three principle forms of tPA in plasma: active tPA, tPA complexed with plasminogen activator inhibitor type 1 (PAI-1), and tPA complexed with C1-inhibitor. METHODS:Active tPA was measured by use of an indirect amidolytic as...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2000-01-01 00:00:00
abstract：BACKGROUND:Hepcidin-25 reduces iron absorption by binding to the intestinal iron transporter ferroportin and causing its degradation. Currently, little is known about the basal regulation of circulating hepcidin-25. In addition, although erythropoietin administration has been reported to decrease the circulating hepcid...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.186866
更新日期：2012-08-01 00:00:00
abstract：:We describe a procedure based on equilibrium dialysis that allows the simultaneous determination of free testosterone and testosterone bound to non-sex-hormone-binding globulin (non-SHBG) in plasma. After saturating SHBG with 5 alpha-dihydrotestosterone (DHT) according to a technique recently described, the percentage...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-06-01 00:00:00
abstract：:We compared results obtained with a commercial immunoradiometric assay kit for human thyrotropin in which monoclonal antibodies are used (Tandem -R TSH (ONE STEP) ImmunoRadioMetricAssay; Hybritech Inc.) with those obtained with two commercial radioimmunoassay methods: GAMMA-DAB [125I]HS-hTSH RIA (Clinical Assays) and ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-06-01 00:00:00
abstract：:In this method for measuring amino acids in urine, serum, and tissue, the amino acids to be assayed and the internal standard, L-norleucine, are converted to phenylthiohydantoins , isolated by organic solvent extraction, separated by reversed-phase "high-pressure" liquid chromatography, and detected by ultraviolet abs...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-06-01 00:00:00
abstract：:We have studied the influence of triglyceride-rich particles on the analytical bias of apolipoprotein B measurements by various immunoturbidimetric methods. Three commercially available methods grossly overestimate apolipoprotein B in samples with even moderately above-normal triglyceride concentrations. This effect i...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-05-01 00:00:00
abstract：:We describe a rapid, reliable radioimmunoassay for unconjugated estriol in plasma. Polyethylene glycol (Carbowax 6000) is used to separate antibody-bound and free steroid. The assay is sensitive (25 pg for standards), precise, and accurate. At high and low concentrations of estriol, intra-assay coefficients of variati...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1976-03-01 00:00:00
abstract：:Investigating the feasibility and validity of determining plasma ferritin concentration in blood obtained by finger prick, we studied 29 adults (ages 21-49 years) and 35 children (ages 14-66 months). Blood was sampled simultaneously in the same subject from both the antecubital vein (venous blood) and by finger pricki...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-05-01 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
