Applications

The most important determinant for the suitability for tissue transplantation has been the HI.A type of the individuals. Traditionally, HLA types have been

= template

= newly-synthesized DNA

Figure 2 The principles of PCR amplification.

= template

= newly-synthesized DNA

Figure 2 The principles of PCR amplification.

identified by the use of antibodies. Now it is common to use PCR; this procedure is faster and more accurate. Specific primers are used to amplify a number of HLA genes in a single tube. After the amplification procedure the products are analyzed by gel electrophoresis. The degree of similarity (and therefore compatibility) is indicated by the banding pattern of the donor and the recipient.

Within the human genome are a number of loci known as microsatellites or short tandem repeats (STRs). As these repeated sequences vary from individual to individual they can be used for genetic mapping. Most often these microsatellites vary in the number of CA nucleotide repeats; these repeats can be detected by using synthetic primers that amplify unique sequences either side of a CA repeat. Different alelle sizes are seen by the migration of amplified DNA on a polyacrylamide or agarose gel. In a standard reaction seven primer pairs are used; these not only have a discriminatory power of greater than 1 X 10"'' but can also be used to confirm the sex of an individual using primer sequences based on the human gene amelogenin, a gene that encodes a protein that makes up part of the tooth enamel.

As PCR procedures become widespread, it may be that many of the antibody-based techniques used in immunology ranging from tissue typing to microbial identification will be replaced by DNA-based methods.

See also: Chromosome translocations of immune genes; Genetic analysis at the molecular level; Northern blotting; Southern blotting; Tissue typing.

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