Real-time Taqman probe technique system for detecting the MtDNA 1555 A>G mutation
10.3760/cma.j.issn.1673-0860.2009.02.019
- VernacularTitle:实时荧光定量Taqman探针法检测线粒体基因1555 A》G突变
- Author:
Yong-Yi YUAN
1
;
De-Liang HUANG
;
Dong-Yi HAN
;
Zheng-Ce JIN
;
Pu DAI
Author Information
1. 解放军总医院
- Keywords:
Hearing loss;
DNA,mitochondrial;
Pnlymerase chain reaction
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2009;44(2):160-164
- CountryChina
- Language:Chinese
-
Abstract:
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, conveniency, 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.
