BACKGROUND: Thromboxane B2 (TxB2) and particularly 11-dehydrothromboxane B2 (11-dTxB2) are widely used as prognostic risk markers of platelet activation in cardiovascular diseases. The main errors in TxB2 and 11-dTxB2 determination include either low concentrations of circulating TxB2 (1 - 2 pg/mL) and 11-dTxB2 (0.9 - 4.3 pg/mL) or rather high transiency (mean TxB2 half-life is approximately 5 minutes) as well as an incorrect pre-analytical phase set up. The aim of this study was to investigate the impact of a widely used purification step on the results of enzyme immunosorbent assay (EIA)--based measurement of the two selected thromboxanes. METHODS: For the purpose of this study, 20 plasma samples (10 healthy donors, 10 patients under treatment with acetylsalicylic acid) were screened for TxB2 and 11-dTxB2 concentrations using commercial competitive EIA kits (Cayman Chemicals, Tallinn, Estonia; Neogen, Lexington, KY, USA) with or without the introduction of the purification procedure. RESULTS: The purification step does not significantly affect the results of EIA measurements of the two of TxA2 metabolites (TxB2, 11-dTxB2) in human plasma. The levels of TxB2 and 11-dTxB2 determined in the plasma samples were not significantly changed (p < 0.05) when the purification step was omitted compared to the purified samples. CONCLUSIONS: This study establishes a protocol allowing for reliable and reproducible plasma TxB2 and 11-dTxB2 EIA measurement for routine basic screening of platelet function.

Division of Clinical Pharmacology Department of Medicine 1 Thomayer University Hospital Prague Czech Republic

000      

00000naa a2200000 a 4500

001      

bmc12022184

003      

CZ-PrNML

005      

20160215095709.0

007      

ta

008      

120806s2012 gw f 000 0#eng||

009      

AR

035    __

$a (PubMed)22372363

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a gw

100    1_

$a Sadílková, Lenka $u Division of Clinical Pharmacology, Department of Medicine I, Thomayer University Hospital, Prague, Czech Republic $7 xx0234438

245    14

$a The purification step is not crucial in EIA measurements of thromboxane B2 and 11-dehydrothromboxane B2 in human plasma / $c L. Sadilkova, Z. Paluch, J. Mottlova, F. Bednar, S. Alusik

520    9_

$a BACKGROUND: Thromboxane B2 (TxB2) and particularly 11-dehydrothromboxane B2 (11-dTxB2) are widely used as prognostic risk markers of platelet activation in cardiovascular diseases. The main errors in TxB2 and 11-dTxB2 determination include either low concentrations of circulating TxB2 (1 - 2 pg/mL) and 11-dTxB2 (0.9 - 4.3 pg/mL) or rather high transiency (mean TxB2 half-life is approximately 5 minutes) as well as an incorrect pre-analytical phase set up. The aim of this study was to investigate the impact of a widely used purification step on the results of enzyme immunosorbent assay (EIA)--based measurement of the two selected thromboxanes. METHODS: For the purpose of this study, 20 plasma samples (10 healthy donors, 10 patients under treatment with acetylsalicylic acid) were screened for TxB2 and 11-dTxB2 concentrations using commercial competitive EIA kits (Cayman Chemicals, Tallinn, Estonia; Neogen, Lexington, KY, USA) with or without the introduction of the purification procedure. RESULTS: The purification step does not significantly affect the results of EIA measurements of the two of TxA2 metabolites (TxB2, 11-dTxB2) in human plasma. The levels of TxB2 and 11-dTxB2 determined in the plasma samples were not significantly changed (p < 0.05) when the purification step was omitted compared to the purified samples. CONCLUSIONS: This study establishes a protocol allowing for reliable and reproducible plasma TxB2 and 11-dTxB2 EIA measurement for routine basic screening of platelet function.

650    _2

$a antiflogistika nesteroidní $x farmakologie $7 D000894

650    _2

$a Aspirin $x farmakologie $7 D001241

650    _2

$a trombocyty $x účinky léků $7 D001792

650    _2

$a kardiovaskulární nemoci $x krev $x diagnóza $7 D002318

650    _2

$a lidé $7 D006801

650    _2

$a imunoenzymatické techniky $x metody $7 D007124

650    _2

$a aktivace trombocytů $x účinky léků $7 D015539

650    _2

$a prognóza $7 D011379

650    _2

$a reagenční diagnostické soupravy $7 D011933

650    _2

$a reprodukovatelnost výsledků $7 D015203

650    _2

$a extrakce na pevné fázi $x metody $7 D052616

650    _2

$a thromboxan B2 $x analogy a deriváty $x krev $7 D013929

655    _2

$a časopisecké články $7 D016428

700    1_

$a Paluch, Zoltán $7 xx0089928

700    1_

$a Mottlováa, Jiřina

700    1_

$a Bednář, František $7 xx0098618

700    1_

$a Alušík, Štefan, $d 1947- $7 mzk2002160778

773    0_

$w MED00009480 $t Clinical laboratory $x 1433-6510 $g Roč. 58, č. 1-2 (2012), s. 177-183

856    41

$u https://pubmed.ncbi.nlm.nih.gov/22372363 $y Pubmed

910    __

$a ABA008 $b sig $c sign $y m $z 0

990    __

$a 20120806 $b ABA008

991    __

$a 20160215095851 $b ABA008

999    __

$a ok $b bmc $g 944097 $s 779481

BAS    __

$a 3

BAS    __

$a PreBMC

BMC    __

$a 2012 $b 58 $c 1-2 $d 177-183 $i 1433-6510 $m Clinical laboratory $n Clin Lab $x MED00009480

LZP    __

$b NLK111 $a Pubmed-20120806/12/01