|
| Method | Typing principle | Advantages | Disadvantages |
|
Phenotyping
[28–31] | Structure and size
variations in proteins | Used for a long time
Large amount of data accumulated
Detects rare and/or new variants | Cannot detect genotype
Requires special equipment and trained
personnel |
Southern blotting
[16, 20] | Restriction size variation | Detects allele
May recognize new alleles | Labor and timeconsuming
Requires large amount of DNA
Risk of radiation hazard |
Conventional PCR
[13–15, 18] | Size variation of amplified
products | Distinguishes between , ,
and alleles
under appropriate combinations | Need to keep multiple primer sets
Tedious postamplification process
Difficult to amplify and detect
large-sized products |
| Real-time PCR using TaqMan probe [32, 33] | Signals from probes
reacting to amplified
regions and their ratios | Discriminates between , ,
and alleles in a single reaction
| Cannot detect rare variants
Multiple sets of primers and probes
Reaction failure in a large scale study |
| Real-time PCR using SYBR Green I [34] | Melting curve analysis | Detect allele effectively | Cannot distinguish between and
Reaction failure in a large scale study |
Loop-mediated isothermal amplification
[35] | Turbidity measurement | Detect allele effectively
No need for a thermal cycler | Cannot distinguish between and
Multiple sets of primers and 2 reaction tubes needed
Not thoroughly evaluated |
|
