title: HIV contamination of commercial PCR enzymes raises the importance of quality control of low-cost in-house genotypic HIV drug resistance tests.
abstract: BACKGROUND
  Low-cost in-house technologies for genotypic drug resistance testing use reagents with quality labels for research only. Here, we report on the results of PCR amplifications in negative-controls that were observed in two independent laboratories.
  
  
  METHODS
  Positive PCR amplifications of protease and reverse transcriptase fragments for genotypic drug resistance testing of HIV on dried blood and/or plasma spots were observed on negative-control samples and were analysed in detail by PCR and sequence and phylogenetic analyses to identify the origin of the PCR contamination.
  
  
  RESULTS
  Detailed analysis revealed that the RT-PCR enzymes were contaminated with an HIV-based vector commercialized by the same company.
  
  
  CONCLUSIONS
  These observations show the need to implement quality control steps that verify for the absence of HIV in new reagent batches because this can significantly compromise molecular diagnosis of HIV and genotypic drug resistance tests using in-house protocols.
authors.affiliation: UMR 145, Institut de Recherche pour le Développement, Université de Montpellier 1, Montpellier, France.
authors.forename: Marjorie
authors.initials: M
authors.lastname: Monleau
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Plantier
authors.forename: Martine
authors.initials: M
authors.lastname: Peeters
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D024882:Drug Resistance, Viral; D004866:Equipment Contamination; D005189:False Positive Reactions; D005822:Genetic Vectors; D015658:HIV Infections / Q000175:diagnosis / Q000821:virology; D016333:HIV Protease / Q000032:analysis / Q000235:genetics; D054303:HIV Reverse Transcriptase / Q000032:analysis / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D011786:Quality Control; D011933:Reagent Kits, Diagnostic / Q000191:economics / Q000592:standards*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000592:standards*; D017422:Sequence Analysis, DNA
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D054303:HIV Reverse Transcriptase; D016333:HIV Protease