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Updated: Jun 22, 2026
Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
Published on: February 10, 2023
[Real-time Taqman probe technique system for detecting the MtDNA 1555 A > G mutation.]
Yong-Yi Yuan1, De-Liang Huang, Dong-Yi Han
1Department of Otorhinolaryngology Head and Neck Surgery, Institute of Otorhinolaryngol, Genetic Testing Center for Deafness, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Objective:
To establish a Real-time Taqman probe technique system to detect the mtDNA 1555A > G mutation in deaf population.
Methods:
Primers and Taqman probes for mtDNA 1555A > G mutation were designed and synthesized. The technique system for detecting mtDNA 1555A > G mutation using Real-time Taqman probes was established. Then the reliability of the technique was tested in 132 patients with severe to profound hearing loss who were detected for the mtDNA 1555A > G mutation by sequencing, Kit method and Real-time Taqman probe technique at the same time. Finally, the results by the above three ways were compared.
Results:
Thirty-two cases with mtDNA 1555A > G mutation were found by the technique of Real-time Taqman probe. These findings coincided with the results from sequencing and Kit method completely. Both the false positive rate and the false negative rate were zero.
Conclusions:
The technique possesses the merits of accuracy, convenience, high sensitivity, high specificity and intuitionistic results, etc. Importantly, the Real-time Taqman probe technique only needs 1.5 hours to detect the 1555A > G mutation and it saves 4.5 hours for one reaction compared with the Kit method popularly used nowadays. The technique system of detecting mtDNA 1555A > G mutation is reliable. It's suitable for large-scale detecting and preventive diagnosis of mtDNA 1555A > G mutation.
