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

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(2). 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.