Validation of quantitative polymerase chain reaction with Southern blot method for telomere length analysis

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/m. 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.