title: Detection of HLA-B*57:01 by real-time PCR: implementation into routine clinical practice and additional validation data.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute of Pharmacology, Catholic University Medical School, L.go F Vito 1, 00168 Rome, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Dello Russo
authors.forename: Lucia
authors.initials: L
authors.lastname: Lisi
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Fabbiani
authors.forename: Dimitri
authors.initials: D
authors.lastname: Gagliardi
authors.forename: Iuri
authors.initials: I
authors.lastname: Fanti
authors.forename: Simona
authors.initials: S
authors.lastname: Di Giambenedetto
authors.forename: Roberto
authors.initials: R
authors.lastname: Cauda
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Navarra
mesh_terms: D000328:Adult; D015224:Dideoxynucleosides / Q000008:administration & dosage*; D004342:Drug Hypersensitivity; D005260:Female; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy / Q000235:genetics*; D015235:HLA-B Antigens / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D010597:Pharmacogenetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D000099227:Abacavir
chemical_list: D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D000099227:Abacavir; C484830:HLA-B*57:01 antigen