In case that the mutation leading to the Hemophilia A or B is not known or the direct mutation detection is not possible, the indirect diagnosis or heteroduplex analysis
is alternatives for the carrier detection in families at risk (Wulff et al.
Applications of heteroduplex analysis
for mutation detection in disease genes.
They noted a higher positivity rate with TGGE-based analysis compared to heteroduplex analysis
and sequencing gel analysis.
The PCR products were subjected to mutation screening by single-strand conformation polymorphism and heteroduplex analysis
This form of heteroduplex analysis
has focused on detection, and although it may involve a quantitative threshold, the result is essentially binary.
Earlier studies on heteroduplex analysis
suggested that the kinetics of heteroduplex formation would fit a quadratic model (26, 27).
Capillary and microchip electrophoresis for rapid detection of known mutations by combining allele-specific DNA amplification with heteroduplex analysis
In these cases, there are 2 options to retain the simplicity of closed-tube genotyping with a dye: (a) wild-type DNA can be added to all samples and quantitative heteroduplex analysis
performed (20); or (b) an unlabeled probe can be included in the PCR to differentiate the homozygotes (28).
Several approaches have been developed for the analysis of cancer susceptibility genes by microchip CE, including single-strand conformation polymorphism analysis (33) and a combination of allele-specific DNA amplification with heteroduplex analysis
These methods include single-strand conformation polymorphism analysis, denaturing gradient gel electrophoresis, temperature gradient gel electrophoresis, heteroduplex analysis
, denaturing HPLC, temperature gradient capillary electrophoresis, and chemical or enzymatic mismatch detection methods (25).
Other genotyping methods could also be affected by adjacent sequence alterations, including allele-specific amplification (6), single-strand conformational polymorphism analysis (7), oligonucleotide ligation (8), heteroduplex analysis
(9), and methods based on melting curve analysis, in which unexpected results should be clarified definitively by sequencing (10).
Indeed, using heteroduplex analysis
and subsequently verifying the identified changes by DNA sequencing, the authors discovered APOE sequence variants in 3 of 765 patients, a frequency that was considerably higher than in nondiabetic patients.