Rapid RHD Zygosity Determination Using Digital PCR.

Abstract:

BACKGROUND:Paternal zygosity testing is used for determining homo- or hemizygosity of RHD in pregnancies that are at a risk of hemolytic disease of the fetus and newborn. At present, this is achieved by using real-time PCR or the Rhesus box PCR, which can be difficult to interpret and unreliable, particularly for black African populations. METHODS:DNA samples extracted from 53 blood donors were analyzed using 2 multiplex reactions for RHD-specific targets against a reference (AGO1)2 to determine gene dosage by digital PCR. Results were compared with serological data, and the correct genotype for 2 discordant results was determined by long-range PCR (LR-PCR), next-generation sequencing, and conventional Sanger sequencing. RESULTS:The results showed clear and reliable determination of RHD zygosity using digital PCR and revealed that 4 samples did not match the serologically predicted genotype. Sanger sequencing and long-range PCR followed by next-generation sequencing revealed that the correct genotypes for samples 729M and 351D, which were serologically typed as R1R2 (DCe/DcE), were R2r' (DcE/dCe) for 729M and R1r″ (DCe/dcE), R0ry (Dce/dCE), or RZr (DCE/dce) for 351D, in concordance with the digital PCR data. CONCLUSIONS:Digital PCR provides a highly accurate method to rapidly define blood group zygosity and has clinical application in the analysis of Rh phenotyped or genotyped samples. The vast majority of current blood group genotyping platforms are not designed to define zygosity, and thus, this technique may be used to define paternal RH zygosity in pregnancies that are at a risk of hemolytic disease of the fetus and newborn and can distinguish between homo- and hemizygous RHD-positive individuals.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Sillence KA,Halawani AJ,Tounsi WA,Clarke KA,Kiernan M,Madgett TE,Avent ND

doi

10.1373/clinchem.2016.268698

subject

Has Abstract

pub_date

2017-08-01 00:00:00

pages

1388-1397

issue

8

eissn

0009-9147

issn

1530-8561

pii

clinchem.2016.268698

journal_volume

63

pub_type

杂志文章
  • Increase in dialyzable calcium associated with therapy with lithium.

    abstract::We measured total and dialyzable calcium concentrations in consecutive sera submitted for routine lithium analysis. Of 98 samples from 61 different individuals, six (6.1%) total calcium results and 32 (33%) dialyzable calcium results were above the respective reference intervals. By comparison, when both total and dia...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Toffaletti J,McComb RB,Bowers GN Jr

    更新日期:1979-10-01 00:00:00

  • Nitromethane interferes in assay of creatinine by the Jaffé reaction.

    abstract::A patient who ingested a methanol/nitromethane mixture (model airplane fuel) showed an apparent plasma creatinine concentration of 8.0 mmol/L by the Jaffé reaction when the actual creatinine concentration, as measured by a specific enzymatic method, was 0.09 mmol/L. This effect was due to nitromethane in the plasma. I...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: De Leacy EA,Brown NN,Clague AE

    更新日期:1989-08-01 00:00:00

  • Radioimmunoassay of cortisone in serum, urine, and saliva to assess the status of the cortisol-cortisone shuttle.

    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:

    authors: Morineau G,Boudi A,Barka A,Gourmelen M,Degeilh F,Hardy N,al-Halnak A,Soliman H,Gosling JP,Julien R,Brerault JL,Boudou P,Aubert P,Villette JM,Pruna A,Galons H,Fiet J

    更新日期:1997-08-01 00:00:00

  • Creatine kinase in serum: 6. Inhibition by endogenous polyvalent cations, and effect of chelators on the activity and stability of some assay components.

    abstract::We studied the effects of some chelators on creatine kinase activity. Creatine kinase is competitively inhibited by endogenous polyvalent cations (e.g., calcium, Ki = 4.5 mmol/L); this can be reversed by adding chelators to the reagent, resulting in a mean increase in activity of 1.14-fold at 25 degrees C and 1.18-fol...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Szasz G,Waldenström J,Gruber W

    更新日期:1979-03-01 00:00:00

  • Analgesia and Opioids: A Pharmacogenetics Shortlist for Implementation in Clinical Practice.

    abstract:BACKGROUND:The use of opioids to alleviate pain is complicated by the risk of severe adverse events and the large variability in dose requirements. Pharmacogenetics (PGx) could possibly be used to tailor pain medication based on an individual's genetic background. Many potential genetic markers have been described, and...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:10.1373/clinchem.2016.264986

    authors: Matic M,de Wildt SN,Tibboel D,van Schaik RHN

    更新日期:2017-07-01 00:00:00

  • Improved measurement of acetylsalicylic acid esterase in serum.

    abstract::A fixed-time incubation method for measuring acetylsalicylic acid esterase was improved by using a higher concentration of the substrate, acetylsalicylic acid, and an activator, Ca2+. The enzymatic activity in serum was 25-fold that measured by earlier methods. Inhibition studies showed a pattern similar to that repor...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Sørensen S

    更新日期:1983-03-01 00:00:00

  • Improved detection of oligoclonal IgG in cerebrospinal fluid by isoelectric focusing in agarose, double-antibody peroxidase labeling, and avidin-biotin amplification.

    abstract::To demonstrate oligoclonal IgG bands (I) in unconcentrated cerebrospinal fluid, we used isoelectric focusing in agarose followed by protein transfer to cellulose nitrate membrane, double-antibody peroxidase labeling, and avidin-biotin amplification. I can be reliably seen after isoelectric focusing of 5-microL specime...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Olsson T,Kostulas V,Link H

    更新日期:1984-07-01 00:00:00

  • A model for medical decision making and problem solving.

    abstract::Clinicians confront the classical problem of decision making under uncertainty, but a universal procedure by which they deal with this situation, both in diagnosis and therapy, can be defined. This consists in the choice of a specific course of action from available alternatives so as to reduce uncertainty. Formal ana...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Werner M

    更新日期:1995-08-01 00:00:00

  • Sensitive and selective liquid-chromatographic assay of fluoxetine and norfluoxetine in plasma with fluorescence detection after precolumn derivatization.

    abstract::We determined fluoxetine (Prozac) and its major metabolite norfluoxetine in plasma by liquid chromatography with fluorescence detection. After liquid-liquid extraction from 1 mL of plasma, the extract was derivatized at room temperature with dansyl chloride, and the highly fluorescent derivatives were chromatographed ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Suckow RF,Zhang MF,Cooper TB

    更新日期:1992-09-01 00:00:00

  • Overview of the complications of diabetes.

    abstract::Currently, we can make the following generalizations about the major long-term complications of diabetes. First, they occur commonly after 10 to 15 years of diabetes. Second, the complications of renal and retinal disease, but not of atherogenic and neurological disease, appear to be related to the severity of hypergl...

    journal_title:Clinical chemistry

    pub_type: 临床试验,杂志文章,评审

    doi:

    authors: Santiago JV

    更新日期:1986-10-01 00:00:00

  • Bias in sensitivity and specificity caused by data-driven selection of optimal cutoff values: mechanisms, magnitude, and solutions.

    abstract:BACKGROUND:Optimal cutoff values for tests results involving continuous variables are often derived in a data-driven way. This approach, however, may lead to overly optimistic measures of diagnostic accuracy. We evaluated the magnitude of the bias in sensitivity and specificity associated with data-driven selection of ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2007.096032

    authors: Leeflang MM,Moons KG,Reitsma JB,Zwinderman AH

    更新日期:2008-04-01 00:00:00

  • High-performance liquid affinity chromatography: rapid immunoanalysis of transferrin in serum.

    abstract::We describe a new method for quantitatively measuring substances of clinical interest by high-performance liquid affinity chromatography (HPLAC). As a model system we selected analysis for transferrin in human serum with immobilized antibodies in a high-performance liquid chromatographic system. SelectiSpher-10 Activa...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Ohlson S,Gudmundsson BM,Wikström P,Larsson PO

    更新日期:1988-10-01 00:00:00

  • Findings of an external quality assessment scheme for determining aluminum in dialysis fluids and water.

    abstract::The analytical performance of laboratories participating in the dialysis fluids and water aluminum program of the Guildford External Quality Assessment Scheme over the period 1986-1993 has been reviewed. For aluminum concentrations > 3.33 mumol/L in dialysis fluids, the between-laboratory CV has fallen from approximat...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Taylor A,Briggs RJ,Cevik C

    更新日期:1994-08-01 00:00:00

  • Enzymatic determination of anti-NAD+ glycohydrolase in serum.

    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:

    authors: Gella FJ,Viver J,Canalias F,Beleta J,Gonzalez-Sastre F

    更新日期:1993-06-01 00:00:00

  • Glucocorticoid receptors in normal leukocytes: effects of age, gender, season, and plasma cortisol concentrations.

    abstract::We measured glucocorticoid receptors (GR) in mononuclear leukocytes (MNL) isolated from peripheral blood of 145 apparently healthy volunteers (86 men and 59 women). An age-related decrease in the number of GR was suggested between subjects younger than 20 years and elderly subjects; there was no apparent seasonal vari...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Tanaka H,Akama H,Ichikawa Y,Homma M,Oshima H

    更新日期:1991-10-01 00:00:00

  • Serum creatine kinase isoenzyme BB is a poor index to the size of various brain lesions.

    abstract::We divided patients with brain lesions into three groups: (a) patients with primary or metastatic brain cancer, (b) brain infarctions, and (c) brain contusion(s). We analyzed each patient's sera for creatine kinase isoenzyme BB (CK-BB), using a monoclonal antibody kit (Impres-BB; International Immunoassay Laboratories...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Schwartz JG,Bazan C 3rd,Gage CL,Prihoda TJ,Gillham SL

    更新日期:1989-04-01 00:00:00

  • No influence of the MDR-1 C3435T polymorphism or a CYP3A4 promoter polymorphism (CYP3A4-V allele) on dose-adjusted cyclosporin A trough concentrations or rejection incidence in stable renal transplant recipients.

    abstract:BACKGROUND:A substantial proportion of the variability in the absorption and clearance of cyclosporin A (CsA) after oral administration has been attributed to variability in liver cytochrome P-450 3A4 (CYP3A4) activity and intestinal P-glycoprotein (P-gp) concentration. A polymorphism in the CYP3A4 promoter region, ter...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: von Ahsen N,Richter M,Grupp C,Ringe B,Oellerich M,Armstrong VW

    更新日期:2001-06-01 00:00:00

  • Precision of sensitivity estimations in diagnostic test evaluations. Power functions for comparisons of sensitivities of two tests.

    abstract::The precision of estimates of the sensitivity of diagnostic tests is evaluated. "Sensitivity" is defined as the fraction of diseased subjects with test values exceeding the 0.975-fractile of the distribution of control values. An estimate of the sensitivity is subject to sample variation because of variation of both c...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Linnet K

    更新日期:1985-04-01 00:00:00

  • Trace metal analysis by laser-enhanced ionization in flames.

    abstract::We review atomic flame spectrometry of metals based on the selective laser-enhancement of the rate of atomic ionization. A discussion of basic principles, prior literature, and instrumentation is followed by an overview of analytical figures of merit for the technique, and a look at its present status and areas for fu...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Travis JC,Turk GC,DeVoe JR

    更新日期:1983-09-01 00:00:00

  • Multianalyte microspot immunoassay--microanalytical "compact disk" of the future.

    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:

    authors: Ekins RP,Chu FW

    更新日期:1991-11-01 00:00:00

  • Acyclic adenine nucleoside phosphonates in plasma determined by high-performance liquid chromatography with fluorescence detection.

    abstract::9-(2-Phosphonylmethoxyethyl)adenine (PMEA), (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine (HPMPA), and (R,S)-9-(3-fluoro-2-phosphonylmethoxypropyl)adenine (FPMPA) are selective antiviral agents with activity against a broad spectrum of DNA viruses and retroviruses. A highly sensitive method has been developed to ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Naesens L,Balzarini J,De Clercq E

    更新日期:1992-04-01 00:00:00

  • Protease inhibitor-associated dyslipidemia in HIV-infected patients is strongly influenced by the APOA5-1131T->C gene variation.

    abstract:BACKGROUND:Hyperlipidemia associated with the protease inhibitor (PI) component of highly active antiretrovial treatment can lead to accelerated atherosclerosis. The apolipoprotein A-V (APOA5) gene, which affects VLDL production and lipolysis, may play a role in PI-induced hyperlipidemia, particularly in individuals wi...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2006.069583

    authors: Guardiola M,Ferré R,Salazar J,Alonso-Villaverde C,Coll B,Parra S,Masana L,Ribalta J

    更新日期:2006-10-01 00:00:00

  • Energy-dispersive x-ray fluorescence spectroscopy and inductively coupled plasma emission spectrometry evaluated for multielement analysis in complex biological matrices.

    abstract::Energy-dispersive x-ray spectroscopy and inductively coupled plasma emission spectrometry were evaluated as methods for routine multielement analysis of biological material. Standard samples included Standard Reference Materials (National Bureau of Standards), compounded mixtures, and supplements that provided a wide ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Irons RD,Schenk EA,Giauque RD

    更新日期:1976-12-01 00:00:00

  • Identification of chromosomally integrated human herpesvirus 6 by droplet digital PCR.

    abstract:BACKGROUND:Human herpesvirus 6 (HHV-6) latently infects a majority of adults. In about 1% of the population HHV-6 exists in a chromosomally integrated form (ciHHV-6) that resides in every somatic and germ cell and can be transmitted through the germ line. Patients with ciHHV-6 have been misdiagnosed and unnecessarily t...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2013.217240

    authors: Sedlak RH,Cook L,Huang ML,Magaret A,Zerr DM,Boeckh M,Jerome KR

    更新日期:2014-05-01 00:00:00

  • Determination of acid alpha-glucosidase protein: evaluation as a screening marker for Pompe disease and other lysosomal storage disorders.

    abstract:BACKGROUND:In recent years, there have been significant advances in the development of enzyme replacement and other therapies for lysosomal storage disorders (LSDs). Early diagnosis, before the onset of irreversible pathology, has been demonstrated to be critical for maximum efficacy of current and proposed therapies. ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Umapathysivam K,Whittle AM,Ranieri E,Bindloss C,Ravenscroft EM,van Diggelen OP,Hopwood JJ,Meikle PJ

    更新日期:2000-09-01 00:00:00

  • Avidin-biotin enzyme immunoassay of osteocalcin in serum or plasma.

    abstract::We describe a competitive enzyme immunoassay, the ExtrAvidin-biotin system, for determining osteocalcin in human serum or plasma. Antibodies were raised against bovine osteocalcin. Binding of the antibodies to osteocalcin was calcium-dependent. Limit of detection is 0.07 nmol/L (0.4 microgram/L). The standard curve fo...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Jaouhari J,Schiele F,Dragacci S,Tarallo P,Siest JP,Henny J,Siest G

    更新日期:1992-10-01 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

  • gamma-Glutamyltransferase as a predictor of chronic kidney disease in nonhypertensive and nondiabetic Korean men.

    abstract:BACKGROUND:Little research has been done to examine whether gamma-glutamyltransferase (GGT) is prospectively associated with the development of chronic kidney disease (CKD). We performed a prospective study to examine the association between GGT and the risk for the development of CKD. METHODS:The study cohort include...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2006.078980

    authors: Ryu S,Chang Y,Kim DI,Kim WS,Suh BS

    更新日期:2007-01-01 00:00:00

  • Surface modification with protein A for uniform binding of monoclonal antibodies.

    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:

    authors: Schramm W,Yang T,Midgley AR

    更新日期:1987-08-01 00:00:00

  • Effect of lyophilization on results of five enzymatic methods for cholesterol.

    abstract::We studied the effect of lyophilization of serum pools on the determination of cholesterol with the enzymatic methods used in five automated analyzers: SMAC (Technicon), the RA-1000 (Technicon), the aca (Du Pont), the TDx (Abbott), and the Ektachem 700P (Kodak). We prepared two serum pools: pool A (2.2 g/L) and pool B...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Kroll MH,Chesler R,Elin RJ

    更新日期:1989-07-01 00:00:00