Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine values determined by a modified ELISA improves agreement with HPLC-MS/MS
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
24120947
DOI
10.1016/j.bbrc.2013.09.133
PII: S0006-291X(13)01638-0
Knihovny.cz E-resources
- Keywords
- 8-OxodG, Discrepancy, ELISA, HPLC–MS /MS, Oxidative stress, Urine,
- MeSH
- 8-Hydroxy-2'-Deoxyguanosine MeSH
- Biomarkers urine MeSH
- Deoxyguanosine analogs & derivatives urine MeSH
- Adult MeSH
- Enzyme-Linked Immunosorbent Assay standards MeSH
- Solid Phase Extraction MeSH
- Middle Aged MeSH
- Humans MeSH
- Oxidative Stress MeSH
- Sensitivity and Specificity MeSH
- Tandem Mass Spectrometry standards MeSH
- Chromatography, High Pressure Liquid standards MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 8-Hydroxy-2'-Deoxyguanosine MeSH
- Biomarkers MeSH
- Deoxyguanosine MeSH
ELISA is widely used for urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) analysis. It is the method of choice for laboratories that lack specialized chromatographic instrumentation. It allows fast, high-throughput sample analysis without a need for extensive samples processing. However, a lack of agreement between ELISA and chromatographic methods confines its application to the assessment of relative urinary 8-oxodG levels. We investigated various ELISA modifications, seeking optimal conditions that would yield a good agreement between ELISA and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS /MS). Purification of urine by solid phase extraction (SPE), then incubation with the anti-8-oxodG antibody at 4 °C overnight and subsequent normalization of 8-oxodG levels per urinary creatinine resulted in a near-perfect correlation and agreement in mean levels between ELISA and HPLC-MS /MS (r=0.917, p<0.001; and paired t-test p=0.803, respectively). Our data show that, after introduction of a simple modification, ELISA quantification urinary 8-oxodG substantially improves. Although more sample manipulation is required, the method retains its key advantages over chromatography (high-throughput analysis that does not require expensive instrumentation). This represents a significant advance for the ELISA, and encouraging its use in more studies adding to our knowledge of the role of this biomarker of oxidative stress in health and disease.
References provided by Crossref.org
Biomarkers of nucleic acid oxidation - A summary state-of-the-art
