Abstract:
BACKGROUND:The time required for bloodstream pathogen detection, identification (ID), and antimicrobial susceptibility testing (AST) does not satisfy the acute needs of disease management. Conventional methods take up to 3 days for ID and AST. Molecular diagnostics have reduced times for ID, but their promise to supplant culture is unmet because AST times remain slow. We developed a combined quantitative PCR (qPCR)-based ID+AST assay with sequential detection, ID, and AST of leading nosocomial bacterial pathogens. METHODS:ID+AST was performed on whole blood samples by (a) removing blood cells, (b) brief bacterial enrichment, (c) bacterial detection and ID, and (d) species-specific antimicrobial treatment. Broad-spectrum qPCR of the internal transcribed spacer between the 16S and 23S was amplified for detection. High-resolution melting identified the species with a curve classifier. AST was enabled by Ct differences between treated and untreated samples. RESULTS:A detection limit of 1 CFU/mL was achieved for Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. All species were accurately identified by unique melting curves. Antimicrobial minimum inhibitory concentrations were identified with Ct differences of ≥1 cycle. Using an RNA target allowed reduction of AST incubation time from 60 min to 5 min. Rapid-cycle amplification reduced qPCR times by 83% to 30 min. CONCLUSIONS:Combined, sequential ID+AST protocols allow rapid and reliable detection, ID, and AST for the diagnosis of bloodstream infections, enabling conversion of empiric to targeted therapy by the second dose of antimicrobials.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Andini N,Hu A,Zhou L,Cogill S,Wang TH,Wittwer CT,Yang Sdoi
10.1373/clinchem.2018.290189subject
Has Abstractpub_date
2018-10-01 00:00:00pages
1453-1462issue
10eissn
0009-9147issn
1530-8561pii
clinchem.2018.290189journal_volume
64pub_type
杂志文章abstract:BACKGROUND:Platelet-derived microparticles (PMPs) are generally considered a marker of platelet activation in cardiovascular disease. We studied the extent to which PMP subpopulations parallel platelet activation in vitro and in vivo. METHODS:Using flow cytometry, we analyzed PMP subpopulations from resting and activa...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:10.1373/clinchem.2005.057414
更新日期:2006-04-01 00:00:00
abstract::Ingestion of oleander plant, containing the cardiac glycoside oleandrin, has been reported to induce fatal poisonings. Derivatives of oleandrin are structurally similar to digoxin. We investigated the cross-reactivities of oleandrin and its aglycone metabolite, oleandrigenin, in several commercially available digoxin ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-10-01 00:00:00
abstract::A radioimmunoassay for barbiturates is shown to be equally sensitive to phenobarbital and its major urinary metabolite, p-hydroxyphenobarbital, in serum and urine. Interference by the metabolite can be essentially eliminated by selectively extracting phenobarbital into chloroform. The extraction efficiency of the meth...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1977-05-01 00:00:00
abstract::This new one-step chemiluminescent immunoassay of free thyroxin (FT4) involving a thyroxin-immunoglobulin conjugate labeled with acridinium ester (Magic Lite System; Ciba Corning Diagnostics Corp., Medfield, MA) is rapid (one 1-h incubation), requires two calibrators per run, and takes 10 s per sample for the quantifi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-12-01 00:00:00
abstract::We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic pro...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
abstract::We compared a stabilized-temperature L'vov platform furnace containing an end-heated graphite atomizer (HGA) and transverse Zeeman background-correction system with a side-heated furnace system (transversely heated graphite atomizer; THGA) containing a longitudinal Zeeman background-correction system for the determina...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-03-01 00:00:00
abstract:BACKGROUND:The aim of our present study was to establish new reference intervals for thyrotropin (TSH) and thyroid hormones based on National Academy of Clinical Biochemistry (NACB) criteria and regular thyroid ultrasonography. We also assessed the effect of potentially confounding factors to modulate the limits of the...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2004.047399
更新日期:2005-08-01 00:00:00
abstract:BACKGROUND:KLK5 is a newly discovered human kallikrein gene. Many kallikrein genes have been found to be differentially expressed in various malignancies, and prostate-specific antigen (PSA; encoded by the KLK3 gene) is the best tumor marker for prostate cancer. Like the genes that encode PSA and other kallikreins, the...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2002-08-01 00:00:00
abstract:BACKGROUND:The B-type natriuretic peptides (BNP and N-terminal pro-BNP) are secreted by the heart and, in the case of BNP, serve to maintain circulatory homeostasis through renal and vascular actions and oppose many effects of the renin-angiotensin system. Recent evidence suggests that in patients with severe heart fai...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2013.210435
更新日期:2014-03-01 00:00:00
abstract:BACKGROUND:Serial measurements of neurohormones have been shown to improve prognostication in the setting of acute heart failure (HF) or chronic HF without therapeutic intervention. We investigated the prognostic role of serial measurements of emerging neurohormones and BNP in a cohort of chronic HF patients undergoing...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2009.125856
更新日期:2010-01-01 00:00:00
abstract::We describe a sensitive method for the determination of vanadium in biological substances by atomic absorption spectroscopy. By using background correction and pyrolytically coated graphite tubes with flameless apparatus, this vanadium assay has a sensitivity of 65 pg and a detection limit of 30 pg, approximately sixf...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1982-01-01 00:00:00
abstract::We are developing a method for quantifying plasma lipoproteins by proton nuclear magnetic resonance (NMR) spectroscopy that offers advantages over existing clinical methods. We showed that the major lipoproteins have distinct NMR properties sufficient to permit their concentrations to be extracted from a computer line...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-09-01 00:00:00
abstract:BACKGROUND:A sensitive gas chromatography-negative ion chemical ionization mass spectrometry (GC/MS-NICI) method was developed and validated for the measurement of Delta(9)-tetrahydrocannabinol (THC) in human sweat patches. METHODS:THC-d(0) and THC-d(3) were added to worn blank sweat patches (PharmChek; PharmChem Inco...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:10.1373/clinchem.2004.034868
更新日期:2004-11-01 00:00:00
abstract::Proteinuria, long recognized as a sign of renal disease, is a common laboratory abnormality in children and adults. Proteinuria may be transient or persistent; when persistent, the cause must be determined. Proteinuria may be primarily glomerular or tubular in origin. Glomerular proteinuria is usually an important con...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1989-05-01 00:00:00
abstract:BACKGROUND:Previous methods to measure glycohemoglobin (GHb) have been time-consuming or imprecise; we therefore developed a new enzymatic assay for GHb. METHODS:Blood cells were first hemolyzed, and hemoglobin was digested with protease to yield fructosyl amino acid. Fructosyl amino acid oxidase acts on the fructosyl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/49.2.269
更新日期:2003-02-01 00:00:00
abstract::We describe the development of a stable reference material for prostatic acid phosphatase, derived from human prostatic tissue and human seminal fluid. The enzyme was purified by an L-tartramic acid affinity-chromatography technique. Two-dimensional electrophoresis revealed essentially no contaminating proteins, and s...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-08-01 00:00:00
abstract:BACKGROUND:Noninvasive prenatal diagnosis of monogenic disorders using maternal plasma and targeted massively parallel sequencing is being investigated actively. We previously demonstrated that comprehensive genetic diagnosis of a Duchenne muscular dystrophy (DMD) patient is feasible using a single targeted sequencing ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2014.236380
更新日期:2015-06-01 00:00:00
abstract::The increasing use of monoclonal antibodies (MAbs) for disease diagnosis and therapy has created a class of patients at risk for systematic error in clinical testing due to interference by human anti-murine antibodies (HAMA). HAMA interference is often difficult to detect and can cause either an increase or a decrease...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-03-01 00:00:00
abstract::To secure a successful product in today's era of cost containment, manufacturers and developers of new in vitro diagnostic technologies must consider payment issues--coverage and reimbursement by healthcare insurers--as critical market forces that must be addressed in the business plan. Similar to the regulatory hurdl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-09-01 00:00:00
abstract:BACKGROUND:In patient and population samples, generation of analytical results that are comparable and independent of the measurement system, time, and location is essential for the utility of laboratory information supplied in healthcare. Obtaining analytical measurement results with such characteristics is the aim of...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2008.107052
更新日期:2009-06-01 00:00:00
abstract::We present a case of advanced ovarian carcinoma associated with high concentrations of serum and urine amylase. Biochemical analysis to elucidate the nature and origin of the hyperamylasemia identified seven amylase isoenzymes, of which five appeared to be distinct from normal salivary and pancreatic amylase. We discu...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-06-01 00:00:00
abstract::We evaluated the technical performance of the Ciba Corning ACS:180 automated immunoassay system for the following analytes: thyroid-stimulating hormone, free thyroxine, luteinizing hormone, follicle-stimulating hormone, prolactin, human chorionic gonadotropin, carcino-embryonic antigen, and prostate-specific antigen. ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-03-01 00:00:00
abstract::We have devloped an enzyme-linked immunosorbent assay for determining choriomammotropin (human placental lactogen) in serum. Unlabeled hormone competes with choriomammotropin-beta-galactosidase conjugate for antibody bound to polystyrene tubes. The entire assay can be performed in 2.5 h with good precision. The coeffi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-02-01 00:00:00
abstract:BACKGROUND:Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone that exists in multiple forms. Immunoassays commonly used in clinical laboratories measure intact hCG, total beta hCG (intact hCG + hCG free beta-subunit), and/or hCG free beta-subunit. Measurement of serum concentrations of hCG is us...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2007.098822
更新日期:2008-04-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2005.065979
更新日期:2006-08-01 00:00:00
abstract::The clinical value of measuring concentrations of methylmalonic acid in serum (S-MMA) as an aid in the diagnosis of cobalamin deficiency has recently aroused interest. In 58 healthy subjects, ages 40-68 years, we found a 0.95 reference interval of 0.05-0.37 mumol/L (mean 0.21, SD 0.094). In 33 of the subjects, who wer...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-07-01 00:00:00
abstract::We present data on a filter-paper collection and assay method for measurement of glycated hemoglobin (gHb). Onto filter paper dipped into a solution of glucose oxidase (EC 1.1.3.4) and allowed to dry, approximately 20 microL of capillary blood was spotted. For analysis, we eluted the dried blood spot from the paper by...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-05-01 00:00:00
abstract:BACKGROUND:The use of nonstandardized N-terminal pro-B-type natriuretic peptide (NT-proBNP) assays can contribute to the misdiagnosis of heart failure (HF). Moreover, there is yet to be established a common consensus regarding the circulating forms of NT-proBNP being used in current assays. We aimed to characterize and...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2012.200204
更新日期:2013-10-01 00:00:00
abstract:BACKGROUND:Studies of the effects of tobacco smoke often rely on reported exposure to cigarette smoke, a measure that is subject to bias. We describe here the relationship between parental smoking exposure as assessed by urinary cotinine excretion and lung function in children with asthma. METHODS:We studied 90 childr...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:
更新日期:1999-04-01 00:00:00
abstract::Estimation of plasma catecholamine concentrations by "high-performance" liquid chromatography, although increasingly used, still presents difficulties. We describe a method for clean-up of plasma samples on cation-exchange resin before they are extracted with alumina. This procedure eliminates many of the interfering ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-07-01 00:00:00