Urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine analysis by an improved ELISA: An inter-laboratory comparison study

Rössner, Pavel; Orhan, Hilmi; Koppen, Gudrun; Sakai, Kazuo; Santella, Regina M.; Ambroz, Antonin; Rössnerová, Andrea; Srám, Radim J.; Cigánek, Miroslav; Neča, Jiřı́; Arzuk, Ege; Mutlu, Neliye; Cooke, Marcus S.

doi:10.1016/j.freeradbiomed.2016.03.016

Cited by 24 publications

(22 citation statements)

References 24 publications

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“…MS/MS based on a previous report[15]; the authors demonstrated that purification of urine samples by SPE can improve the correlation between these two methods. However, a recent report from this group showed that ELISA still cannot be considered as a robust alternative to HPLC-MS/MS after an inter-laboratory comparison(18]. Our results indicate that the levels of…”

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confidence: 60%

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Comparison of an HPLC-MS/MS Method with Multiple Commercial ELISA Kits on the Determination of Levels of 8-oxo-7,8-Dihydro-2'-Deoxyguanosine in Human Urine

Chen

Zhu

Sun

et al. 2018

JNDC

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Introduction: Analysis of 8-oxodG is usually conducted by either chromatography-based methods or by immunochemical methods commonly used based upon their low cost and high-throughput. However, concern regarding the accuracy of ELISA methods has complicated their use. We directly compare the levels of urinary 8-oxodG obtained by HPLC-MS/MS with three commercially available ELISA kits in this report. Methods: In the current study, a total of 9 human urine samples were analyzed by LC-MS/MS and three commonly used commercial available ELISA kits. Results: We found that urinary 8-oxodG levels analyzed by HPLC-MS/MS [1.4 ± 0.3 nmol/mmol creatinine) were 7.6- to 23.5-fold lower than those detected by ELISA. Overall, the correlations between ELISA and HPLC-MS/MS were poor but were improved after SPE purification for kits from ENZO (P = 0.2817 without SPE; P = 0.0086 with SPE) and Abcam (P = 0.0596 without SPE; P = 0.0473 with SPE). Discussion and conclusion: While we confirmed that SPE purification can improve the correlation between the selected ELISA kits and HPLC-MS/MS, HPLC-MS/MS is still the method of choice to accurately assess the levels of 8-oxodG in human urine.

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confidence: 60%

“…The analysis of 8-oxodG by ELISA were criticized for its inter-laboratory variability and poor agreement with chromatographic techniques [18]; however, its advantages include the capacity for high-throughput and relatively low in cost [15], that can account for its continued common usage.…”

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confidence: 99%

Comparison of an HPLC-MS/MS Method with Multiple Commercial ELISA Kits on the Determination of Levels of 8-oxo-7,8-Dihydro-2'-Deoxyguanosine in Human Urine

Chen

Zhu

Sun

et al. 2018

JNDC

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“…Given these conflicting findings, we believe that readers should pay attention to the following points: First, the nomenclature used is not consistent across authors, and the abbreviations for the same chemical entity differ over time and among authors, which may confuse readers and cause them to draw a different conclusion. Second, various methods have been used to determine the levels of 8-oxodGuo, and historically, the quantitation of urinary 8-oxodGuo in diabetic nephropathy research has mainly been based on HPLC with electrochemical detection (HPLC-ECD) or ELISA [ 5 , 6 , 20 – 22 ], methods that are hampered by insufficient specificity and sensitivity [ 23 ].…”

Section: Discussionmentioning

confidence: 99%

Increased Oxidative Damage of RNA in Early‐Stage Nephropathy in db/db Mice

Wang

Luo

Ping

et al. 2017

Oxidative Medicine and Cellular Longevity

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To evaluate RNA oxidation in the early stage of diabetic nephropathy, we applied an accurate method based on isotope dilution high-performance liquid chromatography-triple quadruple mass spectrometry to analyze the oxidatively generated guanine nucleosides in renal tissue and urine from db/db mice of different ages. We further investigated the relationship between these oxidative stress markers, microalbumin excretion, and histological changes. We found that the levels of 8-oxo-7,8-dihydroguanosine (8-oxoGuo) and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) were increased in the urine and renal tissue of db/db mice and db/db mice with early symptoms of diabetic nephropathy suffered from more extensive oxidative damage than lean littermate control db/m mice. Importantly, in contrast to the findings in db/m mice, the 8-oxoGuo levels in the urine and renal tissue of db/db mice were higher than those of 8-oxodGuo at four weeks. These results indicate that RNA oxidation is more apparent than DNA oxidation in the early stage of diabetic nephropathy. RNA oxidation may provide new insight into the pathogenesis of diabetic nephropathy, and urinary 8-oxoGuo may represent a novel, noninvasive, and easily detected biomarker of diabetic kidney diseases if further study could clarify its source and confirm these results in a large population study.

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“…It may also be pointed out that the measurement of 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxodG) in human fluids including urine, plasma and saliva has been achieved by competitive ELISA using N45.1 monoclonal antibody [110,111]. It was recently confirmed that the immunological detection of 8-oxodG in urine is less quantitative than that achieved by HPLC-ESI-MS/MS due to the presence of interfering contaminants such as D-glucose and D-galactose [112,113]. …”

Section: Measurement Of Oxidatively Generated Damage In Cellular Dnamentioning

confidence: 99%

Formation and repair of oxidatively generated damage in cellular DNA

Cadet

Davies

Medeiros

et al. 2017

Free Radical Biology and Medicine

264

228

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In this review article, emphasis is placed on the critical survey of available data concerning modified nucleobase and 2-deoxyribose products that have been identified in cellular DNA following exposure to a wide variety of oxidizing species and agents including, hydroxyl radical, one-electron oxidants, singlet oxygen, hypochlorous acid and ten-eleven translocation enzymes. In addition, information is provided about the generation of secondary oxidation products of 8-oxo-7,8-dihydroguanine and nucleobase addition products with reactive aldehydes arising from the decomposition of lipid peroxides. It is worth noting that the different classes of oxidatively generated DNA damage that consist of single lesions, intra- and interstrand cross-links were unambiguously assigned and quantitatively detected on the basis of accurate measurements involving in most cases high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. The reported data clearly show that the frequency of DNA lesions generated upon severe oxidizing conditions, including exposure to ionizing radiation is low, at best a few modifications per 106 normal bases. Application of accurate analytical measurement methods has also allowed the determination of repair kinetics of several well-defined lesions in cellular DNA that however concerns so far only a restricted number of cases.

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Urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine analysis by an improved ELISA: An inter-laboratory comparison study

Cited by 24 publications

References 24 publications

Comparison of an HPLC-MS/MS Method with Multiple Commercial ELISA Kits on the Determination of Levels of 8-oxo-7,8-Dihydro-2'-Deoxyguanosine in Human Urine

Comparison of an HPLC-MS/MS Method with Multiple Commercial ELISA Kits on the Determination of Levels of 8-oxo-7,8-Dihydro-2'-Deoxyguanosine in Human Urine

Increased Oxidative Damage of RNA in Early‐Stage Nephropathy in db/db Mice

Formation and repair of oxidatively generated damage in cellular DNA

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