Real-time Taqman probe technique system for detecting the MtDNA 1555 A > G mutation..
- Author:
Yong-Yi YUAN
1
;
De-Liang HUANG
;
Dong-Yi HAN
;
Zheng-Ce JIN
;
Pu DAI
Author Information
- Publication Type:Journal Article
- MeSH: Adolescent; Adult; Child; Child, Preschool; DNA Primers; genetics; DNA, Mitochondrial; genetics; Female; Hearing Loss; diagnosis; genetics; Humans; Male; Point Mutation; Polymerase Chain Reaction; methods; Sensitivity and Specificity; Young Adult
- From: Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2009;44(2):160-164
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a Real-time Taqman probe technique system to detect the mtDNA 1555A > G mutation in deaf population.
METHODSPrimers 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.
RESULTSThirty-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.
CONCLUSIONSThe 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.