[Real-time Taqman probe technique system for detecting the MtDNA 1555 A > G mutation.].
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.
Yuan
Yong-Yi
YY
Department of Otorhinolaryngology Head and Neck Surgery, Institute of Otorhinolaryngol, Genetic Testing Center for Deafness, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Huang
De-Liang
DL
Han
Dong-Yi
DY
Jin
Zheng-Ce
ZC
Dai
Pu
P
D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D034381:Hearing Loss / Q000175:diagnosis* / Q000235:genetics*; D006801:Humans; D008297:Male; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D055815:Young Adult
D017931:DNA Primers; D004272:DNA, Mitochondrial