Počet záznamů: 1
Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine analysis by an improved ELISA: An inter-laboratory comparison study
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SYSNO ASEP 0459609 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine analysis by an improved ELISA: An inter-laboratory comparison study Tvůrce(i) Rössner ml., Pavel (UEM-P) RID, ORCID
Orhan, H. (TR)
Koppen, G. (BE)
Sakai, K. (JP)
Santella, R. M. (US)
Ambrož, Antonín (UEM-P)
Rössnerová, Andrea (UEM-P) RID
Šrám, Radim (UEM-P) RID
Ciganek, M. (CZ)
Neca, J. (CZ)
Arzuk, E. (TR)
Mutlu, N. (TR)
Cooke, M.S. (US)Zdroj.dok. Free Radical Biology and Medicine. - : Elsevier - ISSN 0891-5849
Roč. 95, č. 2016 (2016), s. 169-179Poč.str. 11 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova 8-OxodG ; ELISA ; HPLC-MS/MS Vědní obor RIV DN - Vliv životního prostředí na zdraví CEP GA13-13458S GA ČR - Grantová agentura ČR LO1508 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy GA16-14631S GA ČR - Grantová agentura ČR Institucionální podpora UEM-P - RVO:68378041 UT WOS 000376169800014 EID SCOPUS 84961910380 DOI 10.1016/j.freeradbiomed.2016.03.016 Anotace ELISA is commonly used for the detection of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), a marker of whole body oxidative stress. We performed an inter-laboratory comparison of 8-oxodG by ELISA using standardized experimental conditions, including: sample pre-treatment with solid-phase extraction (SPE), performing analysis using a commercial kit from a single manufacturer and strict temperature control during the assay. We compared the ELISA results with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and performed tentative identification of compounds that may contribute to the discrepancy between both methods. For all but one participating laboratory (Data 1) we observed consistent ELISA results lying mostly within 1SD of the mean 8-oxodG concentration. Mean 8-oxodG levels assessed by ELISA correlated with the data obtained by HPLC-MS/MS (R=0.679, p<0.001). The correlation improved when Data 1 were excluded from the analysis (R=0.749, p<0.001). We identified three outlying urine samples; one with an ELISA 8-oxodG concentration lower, and two with 8-oxodG levels higher, than those measured by HPLC-MS/MS. Omitting these samples further improved inter-methodology agreement (R=0.869, p<0.001). In the outliers with high 8-oxodG estimates various aromatic and heterocyclic compounds were tentatively identified using gas chromatography-mass spectrometry (GC-MS). Application of authentic standards revealed the presence of saccharides as putative interfering substances. In summary, assay standardization improved ELISA inter-laboratory agreement, although some variability is still observed. There are still compounds contributing to overestimation of 8-oxodG by ELISA, but only in some urine samples. Thus, despite significant improvement, ELISA still should not be considered a robust alternative to chromatographic techniques. Pracoviště Ústav experimentální medicíny Kontakt Lenka Koželská, lenka.kozelska@iem.cas.cz, Tel.: 241 062 218, 296 442 218 Rok sběru 2017
Počet záznamů: 1