title: A new approach to safely type for HLA the HIV infected people eligible to abacavir therapy: saliva or buccal swab as reliable DNA sources.
abstract: BACKGROUND
  Increasing the safety in Immunogenetics Labs, in the era of antiretroviral pharmacogenomics, represents an imperative goal. To this purpose, we tested saliva and buccal cells as biological sources of DNA, alternative to peripheral blood, for HLA-B*57:01 genomic typing of HIV positive patients eligible to treatment with abacavir.
  
  
  METHODS
  Blood, saliva and buccal cells of 20 voluntary donors and 20 HIV positive patients were collected. DNA was extracted with a manual commercial kit and an automated platform. Quality and quantity of DNA was evaluated with different procedures. The suitability and reliability of DNAs for HLA-B*57:01 genotyping was checked at low and high resolution level, using PCR-SSP (sequence specific primers PCR), revPCR-SSO (reverse sequence specific oligonucleotides PCR), bead array and SBT (sequence based typing) techniques.
  
  
  RESULTS
  DNA concentrations were qualitatively very good and quantitatively comparable in all the specimens tested with an inferior yield for cotton swabs. Comparing the results of HLA typing with different methodologies, the 100% of reproducibility was achieved.
  
  
  CONCLUSIONS
  The viral load of buccal epithelial cells or saliva is extremely low. Here we demonstrated that the DNA from these alternative sources is appropriate for HLA-B*57:01 typing. We strongly recommend the use of this procedure to increase the safety in the lab when dealing with infectious samples.
authors.affiliation: Servizio di Immunematologia e Medicina Trasfusionale e Centro di Immunologia dei Trapianti, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy. c.badulli@smatteo.pv.it
authors.forename: C
authors.initials: C
authors.lastname: Badulli
authors.forename: I
authors.initials: I
authors.lastname: Sbarsi
authors.forename: D
authors.initials: D
authors.lastname: Di Giorgio
authors.forename: M
authors.initials: M
authors.lastname: Mantovani
authors.forename: R
authors.initials: R
authors.lastname: Maserati
authors.forename: G
authors.initials: G
authors.lastname: Barbarini
authors.forename: L
authors.initials: L
authors.lastname: Salvaneschi
authors.forename: M
authors.initials: M
authors.lastname: Martinetti
mesh_terms: D004247:DNA / Q000032:analysis*; D015224:Dideoxynucleosides / Q000627:therapeutic use*; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy*; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D012463:Saliva / Q000187:drug effects* / Q000821:virology; D000099227:Abacavir
chemical_list: D004247:DNA; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D000099227:Abacavir