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Study on the Detection of TP53-R273H/R273C Gene Mutation by Digital PCR |
ZHANG Jiejie1,2,NIU Chunyan2,WANG Xia2,YANG Jingya1,DONG Lianhua2 |
1. College of Food Sciences and Technology,Shanghai Ocean University,Shanghai 201306,China
2. Center for Advanced Measurement Science,National Institute of Metrology,Beijing 100029,China |
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Abstract TP53 gene is the tumor suppressor gene and commonly used as the targeted detection gene in cancer.R273H and R273C are common hot spot mutations in TP53 gene detection.In order to establish a droplet digital polymerase chain reaction (ddPCR) detection method for these two mutations, two sets of ddPCR methods were designed for the mutation sites, and the specificity of the probe was verified.The annealing temperature and primer probe concentration were optimized.The results showed that the established method had good specificity and repeatability, and the correlation coefficients between ddPCR and weight method were 0.9997 and 0.9996,.The limit of detection of R273H and R273C were both 0.02%, and the limit of quantitation were both 0.05%.The methods can be used to accurately detect TP53-R273H and TP53-R273C gene mutations, providing strong support for clinical application.
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Received: 20 December 2023
Published: 05 September 2024
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