Detection of HLA-B*57:01 by real-time PCR: implementation into routine clinical practice and additional validation data.

Cinzia Dello Russo, Lucia Lisi, Simona Di Giambenedetto, Roberto Cauda, Pierluigi Navarra, Massimiliano Fabbiani, Iuri Fanti, Dimitri Gagliardi

Risultato della ricerca: Contributo in rivistaArticolo in rivistapeer review

16 Citazioni (Scopus)

Abstract

Aim:HLA-B*57:01 status needs to be determined before initiating abacavir therapy. We developed a pharmacogenetic real-time (Q)-PCR screening test using two sets of sequence specific primers. This test has been implemented into routine clinical practice. Materials & methods: HIV-infected patients admitted at our University Hospital were thus genotyped using the above mentioned test. A panel of 80 DNA samples with a known genotype were used to characterize Q-PCR conditions using different master mixes. Results: A total of 353 patients were genotyped, detecting 15 (4.25%) HLA-B*57:01 positive carriers. Among the negative patients, 17.2% were treated with abacavir without any hypersensitivity reaction. Using different Q-PCR master mixes, significantly lower cutoff Ct values were found, thus new analytical settings are provided. Conclusion: The pharmacogenetic test developed in our laboratory for the fast screening of HLA-B*57:01 can be successfully implemented into routine clinical practice. All 16 sequences (including an additional six) currently known for the HLA-B*57:01 allele are detected by sequence specific primers used in this test. The Brilliant II SYBR(®) Green QPCR MM (Stratagene) can safely replace the master mix originally used to develop the test.
Lingua originaleEnglish
pagine (da-a)319-327
Numero di pagine9
RivistaPharmacogenomics
DOI
Stato di pubblicazionePubblicato - 2014

Keywords

  • HIV
  • HLA-B*57:01
  • abacavir
  • fast screening
  • hypersensitivity
  • pharmacogenetic test
  • real-time PCR

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