No Inhibitory Effect of Heparinized Blood on Real-Time PCR Analysis of Thrombophilic Mutations

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F22%3A10464860" target="_blank" >RIV/00179906:_____/22:10464860 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11150/22:10464860

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=WZVfhpOZRU" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=WZVfhpOZRU</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

No Inhibitory Effect of Heparinized Blood on Real-Time PCR Analysis of Thrombophilic Mutations

Popis výsledku v původním jazyce

We compared the efficiency of real-time PCR analysis of FII (c.*97G&gt;A, G20210A) and FV Leiden (c.1601G&gt;A) thrombophilic mutations in the samples obtained from venous blood treated with various anticoagulant agents (EDTA, heparin, and sodium fluoride with potassium oxalate), or from clotted venous blood; one hundred samples of wild-type subjects were tested. Genomic DNA extracts and whole blood specimens modified by 90 °C heating were analysed by real-time PCR analysis; cycle threshold values were subsequently evaluated. Real-time PCR analysis for the FII gene assay performed in DNA extracts from EDTA blood samples revealed a median Ct value of 19.3. Similar Ct values were apparent in the DNA extracts obtained from the heparinized blood and sodium fluoride with potassium oxalatetreated samples: 18.5 and 18.9, respectively. Significantly higher Ct values were found in extracts from clotted blood with medians of 20.6 (tubes with inert separation gel) and 20.5 (tubes without the gel, both P &lt; 0.001). The data on the FV real-time PCR analysis were very comparable to the FII assay. In the modified whole blood, the samples treated with heparin salts showed significantly lower Ct values (P &lt; 0.001) in both assays when compared with the samples with EDTA, sodium fluoride with potassium oxalate, and with the samples with clotted blood. Our results indicate that real-time PCR analyses of thrombophilic mutations were not negatively influenced by the presence of heparin salts in collection tubes. Blood samples with various anticoagulants might be exchangeable for each other when DNA analysis of thrombophilic mutations is required.

Název v anglickém jazyce

No Inhibitory Effect of Heparinized Blood on Real-Time PCR Analysis of Thrombophilic Mutations

Popis výsledku anglicky

We compared the efficiency of real-time PCR analysis of FII (c.*97G&gt;A, G20210A) and FV Leiden (c.1601G&gt;A) thrombophilic mutations in the samples obtained from venous blood treated with various anticoagulant agents (EDTA, heparin, and sodium fluoride with potassium oxalate), or from clotted venous blood; one hundred samples of wild-type subjects were tested. Genomic DNA extracts and whole blood specimens modified by 90 °C heating were analysed by real-time PCR analysis; cycle threshold values were subsequently evaluated. Real-time PCR analysis for the FII gene assay performed in DNA extracts from EDTA blood samples revealed a median Ct value of 19.3. Similar Ct values were apparent in the DNA extracts obtained from the heparinized blood and sodium fluoride with potassium oxalatetreated samples: 18.5 and 18.9, respectively. Significantly higher Ct values were found in extracts from clotted blood with medians of 20.6 (tubes with inert separation gel) and 20.5 (tubes without the gel, both P &lt; 0.001). The data on the FV real-time PCR analysis were very comparable to the FII assay. In the modified whole blood, the samples treated with heparin salts showed significantly lower Ct values (P &lt; 0.001) in both assays when compared with the samples with EDTA, sodium fluoride with potassium oxalate, and with the samples with clotted blood. Our results indicate that real-time PCR analyses of thrombophilic mutations were not negatively influenced by the presence of heparin salts in collection tubes. Blood samples with various anticoagulants might be exchangeable for each other when DNA analysis of thrombophilic mutations is required.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

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OECD FORD obor

30100 - Basic medicine

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2022

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Folia Biologica

ISSN

0015-5500

e-ISSN

2533-7602

Svazek periodika

68

Číslo periodika v rámci svazku

5-6

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

5

Strana od-do

201-205

Kód UT WoS článku

001010557100004

EID výsledku v databázi Scopus

2-s2.0-85160966199