Detection of Mutations in Mycobacteria by PCR-SSCP (Single-Strand Conformation Polymorphism)
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There are a number of techniques available for the detection and characterization of mutations that take advantage of the flexibility and power of polymerase chain reaction (PCR) (Table 1 ). Sequencing remains the gold standard, but it is not very efficient as a screening tool, in particular when evaluating large numbers of samples, or multiple or extended stretches of DNA. f the available techniques, single-strand conformation polymorphism analysis (SSCP) appears as one of the most versatile techniques, amenable to automation, and optimal for screening for known or unknown mutations.
Table 1 Main PCR-Based Strategies for Detection of Mutations
|
Technique |
Relevant reference |
|---|---|
|
Sequencing of PCR products |
(16) |
|
Mutation-specific priming |
(17) |
|
Restriction-enzyme analysts of PCR products |
(18) |
|
Selective ohgonucleotide hybridization |
(19) |
|
Constant denaturmg gel electrophoresis (CDGE) |
(20) |
|
Temperature gradient gel electrophoresis (TGGE) |
(21) |
|
Dtdeoxy tingerprmting |
(22) |
|
Heteroduplex formation analysis |
(23) |
|
Cleavase fragment-length polymorphism (CFLP) |
(24) |
|
Ribonuclease A cleavage of mismatched RNA.DNA duplexes |
(25) |
|
Single-strand conformation polymorphism (SSCP) |
(1) |
