[Methodological evaluation on PCR-DGGE technique in detecting DNA mutation and single nucleotide polymorphism].
OBJECTIVE
To study the efficiency of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in detecting the DNA base mutation and single nucleotide polymorphism (SNP).


METHODS
By DNA sequencing and PCR-DGGE respectively, mutations in the D-loop of the mitochondrial genome were detected in patients with acute leukemia in period of diagnosis and remission, which was regarded as constitutional DNA. The comparison of results was made for evaluating above two methods. Also, the wild type DNA and mutational DNA were mixed at different proportions to mimic the presence of DNA polymorphism. PCR-DGGE was done to study its sensitivity for SNP.


RESULTS
The specificity and sensitivity of PCR-DGGE in detecting base mutation were 100% and 97.1% respectively. And PCR-DGGE could even detect the presence of polymorphism at rate of 0.5%.


CONCLUSIONS
PCR-DGGE has high sensitivity and specificity and can be applied to detection of DNA mutation and SNP, which is better method for checking DNA mutations.
Lu
Xiao-jun
XJ
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Jia
Yong-qian
YQ
Fan
Hong
H
Zhang
Lei
L
Jia
Jin
J
D004252:DNA Mutational Analysis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D054629:Genome, Mitochondrial; D006801:Humans; D007938:Leukemia / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity