Measurement of Oxidative Dna Injury in Humans: Evaluation of a Commercially Available Elisa Assay

Priemé, Helene; Loft, Steffen; Cutler, Richard G.; Poulsen, Henrik Enghusen

doi:10.1533/9780857093059.78

Cited by 13 publications

(6 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As might be expected, mass spectrometric methods demonstrated the greatest accuracy and sensitivity. Although the study showed greater consensus than previously expected (5,11,25,28,29), concern remained over ELISA (8); furthermore, all approaches showed significant variability in intra-technique agreement. In a renewed effort to improve inter-assay agreement, we here present a large scale inter-laboratory exercise with 18 laboratories and common calibrants providing 25 data sets on measurement of 8-oxodG in urine by chromatographic and ELISA based assays.…”

contrasting

confidence: 52%

Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2′-deoxyguanosine

Barregård

Möller

Henriksen

et al. 2013

Antioxidants & Redox Signaling

131

107

View full text Add to dashboard Cite

Aims: Urinary 8-oxo-7,8-dihydro-2¢-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical-and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject-and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs Kronos Science, Phoenix, Arizona. ANTIOXIDANTS & REDOX SIGNALINGVolume 18, Number 18, 2013 ª Mary Ann Liebert, Inc. DOI: 10.1089/ars.2012.4714 2377showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed 'spot' samples showed strong correlations with 24 h excretion (the 'gold' standard) of urinary 8-oxodG (r p 0.67-0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine-or SG-adjusted first morning samples are recommended.

show abstract

contrasting

confidence: 52%

Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2′-deoxyguanosine

Barregård

Möller

Henriksen

et al. 2013

Antioxidants & Redox Signaling

131

107

View full text Add to dashboard Cite

show abstract

“…However, with strict temperature control advocated by Yoshida et al, we achieved levels of 4.12 μg/g creatinine, albeit still 4 times greater than LC-MS/MS analysis (1.03 μg/g creatinine) of the same samples [18,19]. Despite these differences in absolute levels, chromatographic and immunoassay approaches have been shown to correlate significantly [25][26][27], although with some notable exceptions [19,28]. Recent data from our laboratory have led us to conclude that whilst the ELISA is useful, the magnitude of the discrepancy in levels by the two approaches, suggests that the ELISA is, at present, unable to specifically determine Analysis of urinary 8-oxopurines 5 absolute levels of urinary 8-oxodG [19].…”

Section: Introductionmentioning

confidence: 98%

Analysis of urinary 8-oxo-7,8-dihydro-purine-2’-deoxyribonucleosides by LC-MS/MS and improved ELISA

Evans¹,

Singh

Mistry³

et al. 2008

Free Radical Research

View full text Add to dashboard Cite

Non-invasive monitoring of oxidative stress is highly desirable. Urinary 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG) is a biologically relevant and convenient analytical target. However, immunoassays can over-estimate levels of urinary 8-oxodG. Measurement of more than one DNA oxidation product in urine would be advantageous in terms of mechanistic information. Urines samples were analysed for 8-oxodG by solid-phase extraction/LC-MS/MS and ELISA. The solid-phase extraction/LC-MS/MS assay was also applied to the analysis of urinary 7,8-dihydro-8-oxo-2'-deoxyadenosine (8-oxodA). Concurring with previous reports, urinary 8-oxodG measured by ELISA was significantly higher than levels measured by LC-MS/MS. However, apparent improvement in the specificity of the commercially available Japanese Institute for the Control of Ageing (JaICA) ELISA brought mean LC-MS/MS and ELISA measurements of urinary 8-oxodG into agreement. Urinary 8-oxodA was undetectable in all urines, despite efficient recovery by solid phase extraction. Exploitation of the advantages of ELISA may be enhanced by a simple modification to the assay procedure, although chromatographic techniques still remain the 'gold standard' techniques for analysis of urinary 8-oxodG. Urinary 8-oxodA is either not present or below the limit of detection of our instrumentation.

show abstract

“…1.67 pmol/ mol creatinine for ELISA vs. 0.41 pmol/ mol creatinine for LC-MS/MS (Cooke, Singh et al 2006)), but this still has not resolved this problem. Further complicating this issue, there have been instances when the ELISA has been shown not to correlate with chromatographic techniques (Prieme, Loft et al 1996;Cooke, Singh et al 2006); whilst in other reports they do correlate, using HPLC-ECD (Shimoi, Kasai et al 2002;Yoshida, Ogawa et al 2002); and LC-GC/MS (Cooke, Rozalski et al 2006). …”

Section: Introductionmentioning

confidence: 99%

Interlaboratory comparison of methodologies for the measurement of urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine

Cooke¹,

Barregård

Mistry³

et al. 2009

Biomarkers

View full text Add to dashboard Cite

Urinary 8-oxo-7,8-dihydro-2"-deoxyguanosine (8-oxodGuo) is widely used as a marker of oxidative stress. Herein we report the comparison of two, distinct chromatographic assays, with ELISA. Chromatographic assays displayed good agreement (r = 0.89, p < 0.0001), whereas there was markedly worse, albeit still significant, agreement with ELISA (HPLC-GC/MS: r=0.43, HPLC-EC: r=0.56; p<0.0001). Mean values differed significantly between chromatographic assays and ELISA (HPLC-GC/MS: 3.86, HPLC-EC: 4.20, ELISA: 18.70 ng/mg creatinine, p<0.0001). Whilst it is reassuring to note good agreement between chromatographic assay, this study reveals significant short-comings in the ELISA, which bring into question its continued use in its present form.

show abstract

Measurement of Oxidative Dna Injury in Humans: Evaluation of a Commercially Available Elisa Assay

Cited by 13 publications

References 18 publications

Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2′-deoxyguanosine

Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2′-deoxyguanosine

Analysis of urinary 8-oxo-7,8-dihydro-purine-2’-deoxyribonucleosides by LC-MS/MS and improved ELISA

Interlaboratory comparison of methodologies for the measurement of urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine

Contact Info

Product

Resources

About