A "Culture" Shift: Broad Bacterial Detection, Identification, and Antimicrobial Susceptibility Testing Directly from Whole Blood.

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 Chem

journal_title

Clinical chemistry

authors

Andini N,Hu A,Zhou L,Cogill S,Wang TH,Wittwer CT,Yang S

doi

10.1373/clinchem.2018.290189

subject

Has Abstract

pub_date

2018-10-01 00:00:00

pages

1453-1462

issue

10

eissn

0009-9147

issn

1530-8561

pii

clinchem.2018.290189

journal_volume

64

pub_type

杂志文章
  • P-selectin- and CD63-exposing platelet microparticles reflect platelet activation in peripheral arterial disease and myocardial infarction.

    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

    authors: van der Zee PM,Biró E,Ko Y,de Winter RJ,Hack CE,Sturk A,Nieuwland R

    更新日期:2006-04-01 00:00:00

  • Inhibition of Na,K-ATPase by oleandrin and oleandrigenin, and their detection by digoxin immunoassays.

    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:

    authors: Jortani SA,Helm RA,Valdes R Jr

    更新日期:1996-10-01 00:00:00

  • Interference by p-hydroxyphenobarbital in the 125I-radioimmunoassay of serum and urinary phenobarbital.

    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:

    authors: Viswanathan CT,Booker HE,Welling PG

    更新日期:1977-05-01 00:00:00

  • One-step chemiluminescent immunoassay of free thyroxin with acridinium-ester-labeled thyroxin evaluated and compared with a two-step radioimmunoassay.

    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:

    authors: Bounaud JY,Bounaud MP,Begon F

    更新日期:1988-12-01 00:00:00

  • Use of monoclonal and polyclonal antibodies to detect prostatic acid phosphatase by immunoblotting.

    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:

    authors: Patel PC,Aubin L,Côte J

    更新日期:1986-10-01 00:00:00

  • Aluminum determined in plasma and urine by atomic absorption spectroscopy with a transversely heated graphite atomizer furnace.

    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:

    authors: Bradley C,Leung FY

    更新日期:1994-03-01 00:00:00

  • New reference intervals for thyrotropin and thyroid hormones based on National Academy of Clinical Biochemistry criteria and regular ultrasonography of the thyroid.

    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

    authors: Kratzsch J,Fiedler GM,Leichtle A,Brügel M,Buchbinder S,Otto L,Sabri O,Matthes G,Thiery J

    更新日期:2005-08-01 00:00:00

  • Human kallikrein gene 5 (KLK5) expression by quantitative PCR: an independent indicator of poor prognosis in breast cancer.

    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:

    authors: Yousef GM,Scorilas A,Kyriakopoulou LG,Rendl L,Diamandis M,Ponzone R,Biglia N,Giai M,Roagna R,Sismondi P,Diamandis EP

    更新日期:2002-08-01 00:00:00

  • B-type natriuretic peptide forms within the heart, coronary sinus, and peripheral circulation in humans: evidence for degradation before secretion.

    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

    authors: Mahagamasekera PG,Ruygrok PN,Palmer SC,Richards AM,Ansell GS,Nicholls MG,Pemberton CJ,Lewis LK,Yandle TG

    更新日期:2014-03-01 00:00:00

  • Prognostic value of emerging neurohormones in chronic heart failure during optimization of heart failure-specific therapy.

    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

    authors: Neuhold S,Huelsmann M,Strunk G,Struck J,Adlbrecht C,Gouya G,Elhenicky M,Pacher R

    更新日期:2010-01-01 00:00:00

  • More sensitive flameless atomic absorption analysis of vanadium in tissue and serum.

    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:

    authors: Stroop SD,Helinek G,Greene HL

    更新日期:1982-01-01 00:00:00

  • Development of a proton nuclear magnetic resonance spectroscopic method for determining plasma lipoprotein concentrations and subspecies distributions from a single, rapid measurement.

    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:

    authors: Otvos JD,Jeyarajah EJ,Bennett DW,Krauss RM

    更新日期:1992-09-01 00:00:00

  • Validated gas chromatographic-negative ion chemical ionization mass spectrometric method for delta(9)-tetrahydrocannabinol in sweat patches.

    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

    authors: Saito T,Wtsadik A,Scheidweiler KB,Fortner N,Takeichi S,Huestis MA

    更新日期:2004-11-01 00:00:00

  • Current concepts in proteinuria.

    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:

    authors: Waller KV,Ward KM,Mahan JD,Wismatt DK

    更新日期:1989-05-01 00:00:00

  • New enzymatic assay for glycohemoglobin.

    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

    authors: Sakurabayashi I,Watano T,Yonehara S,Ishimaru K,Hirai K,Komori T,Yagi M

    更新日期:2003-02-01 00:00:00

  • Development of a stable reference material for prostatic acid phosphatase.

    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:

    authors: Duncan PH,Van Etten RL,MacNeil ML,Shaw LM

    更新日期:1984-08-01 00:00:00

  • Noninvasive prenatal diagnosis of duchenne muscular dystrophy: comprehensive genetic diagnosis in carrier, proband, and fetus.

    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

    authors: Yoo SK,Lim BC,Byeun J,Hwang H,Kim KJ,Hwang YS,Lee J,Park JS,Lee YS,Namkung J,Park J,Lee S,Shin JY,Seo JS,Kim JI,Chae JH

    更新日期:2015-06-01 00:00:00

  • Modified method for determining carcinoembryonic antigen in the presence of human anti-murine antibodies.

    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:

    authors: Morrissey NE,Quadri SF,Kinders R,Brigham C,Rose S,Blend MJ

    更新日期:1993-03-01 00:00:00

  • A parallel revolution: payment for new in vitro diagnostic technologies.

    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:

    authors: Ivor LP

    更新日期:1996-09-01 00:00:00

  • Traceability in laboratory medicine.

    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

    authors: Vesper HW,Thienpont LM

    更新日期:2009-06-01 00:00:00

  • Hyperamylasemia and ovarian carcinoma.

    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:

    authors: Sandiford JA,Chiknas SG

    更新日期:1979-06-01 00:00:00

  • Evaluation of Ciba Corning ACS:180 automated immunoassay system.

    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:

    authors: Mora-Brugués J,Gascón-Roche N,Rodríguez-Espinosa J,Cortés-Rius M,González-Sastre F

    更新日期:1994-03-01 00:00:00

  • Enzyme-linked immunosorbent choriomammotropin assay.

    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:

    authors: Dittmar D,Monji N,Cid A,Malkus H,Castro A

    更新日期:1979-02-01 00:00:00

  • Are laboratories reporting serum quantitative hCG results correctly?

    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

    authors: Cao ZT,Rej R

    更新日期:2008-04-01 00:00:00

  • Increased plasma concentrations of activin a predict intraventricular hemorrhage in preterm newborns.

    abstract:BACKGROUND:Intraventricular hemorrhage (IVH) is a major cause of neurologic disabilities in preterm newborns. We evaluated the use of plasma activin A concentrations to predict the development of perinatal IVH. METHODS:We measured nucleated erythrocyte (NRBC) counts, plasma activin A, hypoxanthine (Hyp), and xanthine ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2005.065979

    authors: Florio P,Perrone S,Luisi S,Vezzosi P,Longini M,Marzocchi B,Petraglia F,Buonocore G

    更新日期:2006-08-01 00:00:00

  • Methylmalonic acid concentrations in serum of normal subjects: biological variability and effect of oral L-isoleucine loads before and after intramuscular administration of cobalamin.

    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:

    authors: Rasmussen K,Møller J,Ostergaard K,Kristensen MO,Jensen J

    更新日期:1990-07-01 00:00:00

  • Collection of blood on filter paper for measurement of glycated hemoglobin by affinity chromatography.

    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:

    authors: Little RR,McKenzie EM,Wiedmeyer HM,England JD,Goldstein DE

    更新日期:1986-05-01 00:00:00

  • Circulating fragments of N-terminal pro-B-type natriuretic peptides in plasma of heart failure patients.

    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

    authors: Foo JY,Wan Y,Schulz BL,Kostner K,Atherton J,Cooper-White J,Dimeski G,Punyadeera C

    更新日期:2013-10-01 00:00:00

  • Urinary cotinine and exposure to parental smoking in a population of children with asthma.

    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:

    authors: Oddoze C,Dubus JC,Badier M,Thirion X,Pauli AM,Pastor J,Bruguerolle B

    更新日期:1999-04-01 00:00:00

  • Sample preparation with ion-exchange resin before liquid-chromatographic determination of plasma catecholamines.

    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:

    authors: Frayn KN,Maycock PF

    更新日期:1983-07-01 00:00:00