1 Ocak 2016 Dergi makalesi Açık Erişim

Rossner, Pavel, Jr.; Orhan, Hilmi; Koppen, Gudrun; Sakai, Kazuo; Santella, Regina M.; Ambroz, Antonin; Rossnerova, Andrea; Sram, Radim J.; Ciganek, Miroslav; Neca, Jiri; Arzuk, Ege; Mutlu, Neliye; Cooke, Marcus S.

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    "creators": [
      {
        "affiliation": "AS CR, Dept Genet Ecotoxicol, Inst Expt Med, Videnska 1083, Prague 14220, Czech Republic", 
        "name": "Rossner, Pavel, Jr."
      }, 
      {
        "affiliation": "Ege Univ, Dept Toxicol, Fac Pharm, Bornova, Turkey", 
        "name": "Orhan, Hilmi"
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      {
        "affiliation": "Flemish Inst Technol Res VITO, Environm Risk & Hlth Unit, Antwerp, Belgium", 
        "name": "Koppen, Gudrun"
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      {
        "affiliation": "Nikken SEIL Co, Japan Inst Control Aging JaICA, Fukuroi, Shizuoka, Japan", 
        "name": "Sakai, Kazuo"
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      {
        "affiliation": "Columbia Univ, Dept Environm Hlth Sci, Mailman Sch Publ Hlth, New York, NY USA", 
        "name": "Santella, Regina M."
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      {
        "affiliation": "AS CR, Dept Genet Ecotoxicol, Inst Expt Med, Videnska 1083, Prague 14220, Czech Republic", 
        "name": "Ambroz, Antonin"
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      {
        "affiliation": "AS CR, Dept Genet Ecotoxicol, Inst Expt Med, Videnska 1083, Prague 14220, Czech Republic", 
        "name": "Rossnerova, Andrea"
      }, 
      {
        "affiliation": "AS CR, Dept Genet Ecotoxicol, Inst Expt Med, Videnska 1083, Prague 14220, Czech Republic", 
        "name": "Sram, Radim J."
      }, 
      {
        "affiliation": "Vet Res Inst, CS-62132 Brno, Czech Republic", 
        "name": "Ciganek, Miroslav"
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      {
        "affiliation": "Vet Res Inst, CS-62132 Brno, Czech Republic", 
        "name": "Neca, Jiri"
      }, 
      {
        "affiliation": "Ege Univ, Dept Toxicol, Fac Pharm, Bornova, Turkey", 
        "name": "Arzuk, Ege"
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      {
        "affiliation": "Ege Univ, Dept Toxicol, Fac Pharm, Bornova, Turkey", 
        "name": "Mutlu, Neliye"
      }, 
      {
        "affiliation": "Florida Int Univ, Dept Environm & Occupat Hlth, Miami, FL 33199 USA", 
        "name": "Cooke, Marcus S."
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    "description": "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. However, the method has been criticized for high inter-laboratory variability and poor agreement with chromatographic techniques. We performed an inter-laboratory comparison of 8-oxodG assessed in 30 urine samples and a urine spiked with four different concentrations of 8-oxodG by ELISA using standardized experimental conditions, including: sample pretreatment with solid-phase extraction (SPE), performing analysis using a commercial kit from a single manufacturer and strict temperature control during the assay. We further 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 1 SD 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, including D-glucose and D-galactose 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. (C) 2016 Elsevier Inc. All rights reserved.", 
    "doi": "10.1016/j.freeradbiomed.2016.03.016", 
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    "journal": {
      "pages": "169-179", 
      "title": "FREE RADICAL BIOLOGY AND MEDICINE", 
      "volume": "95"
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    "publication_date": "2016-01-01", 
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    "title": "Urinary 8-oxo-7,8-dihydro-2 '-deoxyguanosine analysis by an improved ELISA: An inter-laboratory comparison study"
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