Abstract：
BACKGROUND:Immune repertoire sequencing of the T-cell receptor can identify clonotypes that have expanded as a result of antigen recognition or hematological malignancies. However, current sequencing protocols display limitations with nonuniform amplification and polymerase-induced errors during sequencing. Here, we developed a sequencing method that overcame these issues and applied it to γδ T cells, a cell type that plays a unique role in immunity, autoimmunity, homeostasis of intestine, skin, adipose tissue, and cancer biology. METHODS:The ultrasensitive immune repertoire sequencing method used PCR-introduced unique molecular identifiers. We constructed a 32-panel assay that captured the full diversity of the recombined T-cell receptor delta loci in γδ T cells. The protocol was validated on synthetic reference molecules and blood samples of healthy individuals. RESULTS:The 32-panel assay displayed wide dynamic range, high reproducibility, and analytical sensitivity with single-nucleotide resolution. The method corrected for sequencing-depended quantification bias and polymerase-induced errors and could be applied to both enriched and nonenriched cells. Healthy donors displayed oligoclonal expansion of γδ T cells and similar frequencies of clonotypes were detected in both enrichment and nonenriched samples. CONCLUSIONS:Ultrasensitive immune repertoire sequencing strategy enables quantification of individual and specific clonotypes in a background that can be applied to clinical as well as basic application areas. Our approach is simple, flexible, and can easily be implemented in any molecular laboratory.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Johansson G,Kaltak M,Rîmniceanu C,Singh AK,Lycke J,Malmeström C,Hühn M,Vaarala O,Cardell S,Ståhlberg Adoi
10.1093/clinchem/hvaa159subject
Has Abstractpub_date
2020-09-01 00:00:00pages
1228-1237issue
9eissn
0009-9147issn
1530-8561pii
5894686journal_volume
66pub_type
杂志文章abstract：BACKGROUND:Some of the genetic variants of hemoglobin (Hb) and their chemically modified species are known to affect the measurement of Hb A(1c). The purpose of this study was to characterize Hb species in the blood sample of a 74-year-old German male with an exceptionally low Hb A(1c) value. METHODS:Hemolysates from ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.1.137
更新日期：2003-01-01 00:00:00
abstract：:We describe a discrete automated radioimmunoassay system for determining choriomammotropin (placental lactogen) in human serum. With the present system it can be measured in as many as 37 unknown sera (50 muL) and three quality-control sera, in duplicate, within 1.5 h. The time required for sample preparation, incubat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-01-01 00:00:00
abstract：:In this report on trace-element concentrations (As, Ca, Cd, Cl, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Rb, Sb, Se, Zn) in human heart, liver, kidney, aorta, and rib obtained from 200 autopsied patients, we give special attention to sampling procedure, analysis technique, and various sources of error (autolysis...
journal_title：Clinical chemistry
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doi：
更新日期：1987-11-01 00:00:00
abstract：BACKGROUND:Diagnosis of mitochondrial disorders usually requires a muscle biopsy to examine mitochondrial function. We describe our diagnostic procedure and results for 29 patients with mitochondrial disorders. METHODS:Muscle biopsies were from 43 healthy individuals and 29 patients with defects in one of the oxidativ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.062414
更新日期：2006-05-01 00:00:00
abstract：:We used a previously reported agarose-gel electrophoresis system to study the lipoprotein patterns in patients with type 3 hyperlipoproteinemia. All samples from 20 subjects tested revealed the presence of two bands with beta-lipoprotein mobility, irrespective of quantitative changes in plasma lipids induced by diet o...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-06-01 00:00:00
abstract：:A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-01-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1975-03-01 00:00:00
abstract：:We compared measurements of apoprotein A-I (apoA-I) in plasma and in heparin-MnCl2 supernates analyzed by radial immunodiffusion. The apoA-I values were similar when the samples were fresh [n = 41, mean (SD), mg/L: plasma, 1393.4(349); heparin-MnCl2 supernates, 1364.9(332), p less than 0.01], but were greater than 8% ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-06-01 00:00:00
abstract：:Of 98 patients' specimens examined for alkaline phosphatase (EC 3.1.3.1) isoenzymes by electrophoresis on cellulose acetate membrane after incubation with neuraminidase, 50 showed only a single liver or bone isoenzyme staining band; in 15 of these, the tissue origin of the fraction could not be accurately identified f...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-07-01 00:00:00
abstract：:We measured three renal tubular brush-border enzymes (lactate dehydrogenase, LDH, EC 1.1.1.27; gamma-glutamyltransferase, GGT, EC 2.3.2.2; and alkaline phosphatase, AP, EC 3.1.3.1) in morning urine samples from 48 healthy subjects to check whether different storage times and temperatures could modify enzyme concentrat...
journal_title：Clinical chemistry
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doi：
更新日期：1991-08-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-08-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1985-09-01 00:00:00
abstract：:3-Hydroxy-3-methylglutaric aciduria was detected in a newborn. The progress of the dietary therapy for the disorder was monitored by dual-column "high-performance" liquid chromatography with a computer-controlled photodiode array spectrophotometric detector. This procedure is a quick way to detect and monitor the prog...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-01-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.118497
更新日期：2009-04-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.137471
更新日期：2010-05-01 00:00:00
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journal_title：Clinical chemistry
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doi：10.1373/clinchem.2015.239459
更新日期：2015-11-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-08-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-06-01 00:00:00
abstract：:Solid-phase-based immunoassays have traditionally been plagued by nonspecific binding to the solid phase and by slow reaction kinetics relative to reactants that are free to diffuse in solution. We have developed two novel immunoassays in which the solid phase is generated in situ after the specific binding reaction h...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-09-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-10-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1989-02-01 00:00:00
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doi：
更新日期：1986-02-01 00:00:00
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更新日期：2009-05-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-09-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-10-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-05-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1976-05-01 00:00:00
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journal_title：Clinical chemistry
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doi：10.1373/clinchem.2005.059485
更新日期：2006-03-01 00:00:00
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pub_type： 杂志文章
doi：10.1373/clinchem.2005.065581
更新日期：2006-06-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-06-01 00:00:00
