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Direct PCR: a new pharmacogenetic approach for the inexpensive testing of HLA-B*57:01.

Cascella R, Strafella C, Ragazzo M, Zampatti S, Borgiani P, Gambardella S, Pirazzoli A, Novelli G, Giardina E - Pharmacogenomics J. (2014)

Bottom Line: The amplicons obtained by direct PCR can be easily separated on the agarose gel under ultraviolet.As per our results, the direct PCR represents a good alternative to the traditional methods of HLA-B*57:01 pharmacogenetic test, especially for those laboratories or countries where currently available approaches are often not available or not affordable.Furthermore it is an innovative approach, promoting a personalized, safer and cost-effective therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome 'Tor Vergata', Rome, Italy.

ABSTRACT
One of the most successful applications of pharmacogenetics research is the genetic screening for HLA-B*57:01, strongly associated with an increased risk to develop hypersensitivity reaction in HIV-positive patients following abacavir administration. Taking into consideration the limits of current genotyping methodologies, we have developed and validated (150 buccal swabs) an inexpensive pharmacogenetic approach for HLA-B*57:01 typing. In our assay DNA extraction and amplification are combined in one single step (direct PCR protocol), which is performed directly on the biological sample without the need of extraction and sequencing passages. The amplicons obtained by direct PCR can be easily separated on the agarose gel under ultraviolet. As per our results, the direct PCR represents a good alternative to the traditional methods of HLA-B*57:01 pharmacogenetic test, especially for those laboratories or countries where currently available approaches are often not available or not affordable. Furthermore it is an innovative approach, promoting a personalized, safer and cost-effective therapy.

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Related in: MedlinePlus

Nested PCR. M: ladder 50 bp. CN, negative control.
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fig2: Nested PCR. M: ladder 50 bp. CN, negative control.

Mentions: Figure 2 reports the results of the nested PCR conducted on the four HLA-B*57-positive samples previously found by direct PCR. In this case, the specific amplified regions allow us to distinguish the first three subjects as HLA-B*57:01-positive (94 bp, in the figure ‘B57:01'). The fourth one (in the figure ‘B57+') did not present any amplification of HLA-B*57:01 allele, although it showed to be HLA-B*57-positive.


Direct PCR: a new pharmacogenetic approach for the inexpensive testing of HLA-B*57:01.

Cascella R, Strafella C, Ragazzo M, Zampatti S, Borgiani P, Gambardella S, Pirazzoli A, Novelli G, Giardina E - Pharmacogenomics J. (2014)

Nested PCR. M: ladder 50 bp. CN, negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4381103&req=5

fig2: Nested PCR. M: ladder 50 bp. CN, negative control.
Mentions: Figure 2 reports the results of the nested PCR conducted on the four HLA-B*57-positive samples previously found by direct PCR. In this case, the specific amplified regions allow us to distinguish the first three subjects as HLA-B*57:01-positive (94 bp, in the figure ‘B57:01'). The fourth one (in the figure ‘B57+') did not present any amplification of HLA-B*57:01 allele, although it showed to be HLA-B*57-positive.

Bottom Line: The amplicons obtained by direct PCR can be easily separated on the agarose gel under ultraviolet.As per our results, the direct PCR represents a good alternative to the traditional methods of HLA-B*57:01 pharmacogenetic test, especially for those laboratories or countries where currently available approaches are often not available or not affordable.Furthermore it is an innovative approach, promoting a personalized, safer and cost-effective therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome 'Tor Vergata', Rome, Italy.

ABSTRACT
One of the most successful applications of pharmacogenetics research is the genetic screening for HLA-B*57:01, strongly associated with an increased risk to develop hypersensitivity reaction in HIV-positive patients following abacavir administration. Taking into consideration the limits of current genotyping methodologies, we have developed and validated (150 buccal swabs) an inexpensive pharmacogenetic approach for HLA-B*57:01 typing. In our assay DNA extraction and amplification are combined in one single step (direct PCR protocol), which is performed directly on the biological sample without the need of extraction and sequencing passages. The amplicons obtained by direct PCR can be easily separated on the agarose gel under ultraviolet. As per our results, the direct PCR represents a good alternative to the traditional methods of HLA-B*57:01 pharmacogenetic test, especially for those laboratories or countries where currently available approaches are often not available or not affordable. Furthermore it is an innovative approach, promoting a personalized, safer and cost-effective therapy.

Show MeSH
Related in: MedlinePlus