Detection Of Human Platelet Antigens

Many studies have reported successful molecular detection of various HPAs. These techniques are based upon

GP lib -Illa

GPIb-IX-V

Fig. 1 HPA-1 to -5 on glycoprotein receptors. Three receptor complexes on the platelet membrane with HPA-1 to -5 sites (hatched areas) are shown schematically: the platelet cohesion receptor GP IIb-IIIa, the receptor complex GP Ib-IX-V, and the platelet collagen receptor GP Ia-IIa. The GP Ib-IX-V complex exists on the platelet surface with a suggested stoichiometry of 2:2:2:1 (GP Iba/GP Ibp/ GP IX/GP V).

Fig. 1 HPA-1 to -5 on glycoprotein receptors. Three receptor complexes on the platelet membrane with HPA-1 to -5 sites (hatched areas) are shown schematically: the platelet cohesion receptor GP IIb-IIIa, the receptor complex GP Ib-IX-V, and the platelet collagen receptor GP Ia-IIa. The GP Ib-IX-V complex exists on the platelet surface with a suggested stoichiometry of 2:2:2:1 (GP Iba/GP Ibp/ GP IX/GP V).

conventional PCR where post-PCR steps are required. These conventional techniques will be discussed first and will be followed by discussion of newer techniques not requiring post-PCR steps.

Allele- or Sequence-Specific PCR (SSP-PCR)

Polymerase chain reaction with sequence-specific primers (SSP-PCR) is currently the most widely used technique for HPA genotyping. This approach is based on the principle that a 3' mismatched nucleotide in a primer-template hybrid will prevent extension during PCR, therefore inhibiting DNA amplification. That is, when the 3' nucleotide of a primer is complementary to the se quence at the site of allelic variation, a PCR product will be generated. However, when the 3' nucleotide of the primer is mismatched, the amplification of the template will not occur or will proceed with very low efficiency. For the detection of each HPA allele, two sets of primers are designed, each set containing an allele-specific primer and a common primer. After gel electrophoresis of the PCR product, DNA is stained and analyzed under UV light. The HPA genotype is then determined by the absence or presence of a DNA fragment. As an internal control for the success of the PCR reaction, an additional pair of primers for the amplification of the human growth hormone gene or another gene is usually added to each reaction.

Table 2 Glycoprotein receptors with their subunits and HPAs

GP receptor

Subunit composition

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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