Abstract
Aim:Telomere length (TL) measurement by quantitative polymerase chain reaction (PCR) has been widely accepted, but limited information regarding its validation with a gold-standard technique is available.Materials & methods:We measured TL by Southern blot and monochrome multiplex quantitative PCR (MMqPCR) and validated the results of TL in leukocytes of 94 participants with mean age 43.2 years, BMI 19–41 (mean 27.8 ± 4.3) kg/m2.Results:A significant positive correlation was observed between TL measured by MMqPCR and Southern blot assay (correlation coefficient r = +0.896, p < 0.0001). The inter- and intra-assay CVs of the MMqPCR assay were 5.3 and 4.07%, respectively.Conclusion:We observed that experimental discrepancies have an impact on TL analysis and there is a need to improve the optimum conditions.
Highlights
A strong correlation was observed between both methods and reproducibility of the results from quantitative polymerase chain reaction (qPCR) met acceptance criteria in interassay and intra-assay assessment
Since the sample size was small in the present study, a larger study with more samples spanning different ages so that a wider range of TL is covered would be of interest to assess correlation
Summary
Telomere length (TL) measurement by quantitative polymerase chain reaction (PCR) has been widely accepted, but limited information regarding its validation with a gold-standard technique is available. Lay abstract: The aim of this study was to validate the relative telomere length measurements by different methods. The aim of this study was to validate the relative TL (T/S ratio) measured by MMqPCR by comparing with the gold standard absolute TL measured by Southern blot technique
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