title: Evaluation of NucliSens-AmpliScreen methodology to detect subtypes G of HIV-1 and 4c/4d of HCV in the screening of blood donors.
abstract: OBJECTIVE
  Albeit, the NucliSens Extractor combined with the Ampliscreen was validated for application in NAT minipool screening, a study to evaluate the reliability of the procedure in relation to subtypes G of human immunodeficiency virus (HIV)-1 RNA and 4c/4d of hepatitis C virus (HCV) RNA should be performed, due to their genetic differences and the high frequency in our country.
  
  
  METHODS
  Samples from patients infected with subtypes G of HIV-1 RNA and 4c/4d of HCV RNA were diluted with negative plasma and tested eight times for each concentration. For nucleic acid extraction we used an automated silica-based extraction method (NucliSens Extractor) and for amplification and detection the AmpliScreen HIV-1 version 1.5 and AmpliScreen HCV version 2.0 (Roche Diagnostic Systems) were applied.
  
  
  RESULTS
  The sensitivity for HIV-1 RNA genotype G using the NucliSens-AmpliScreen method-95% detection limit (95% CI) of 25 (18-50) copies per ml-is comparable with those described for genotypes B and E and to that obtained by the Multiprep procedure. In the case of HCV, the sensitivity of the method was also similar, when we compared the detection limits obtained for genotype 4c/4d-95% detection limit (95% CI) of 34 (24-71) IU/ml-with the genotype 1 published.
  
  
  CONCLUSIONS
  The data presented here suggest that these infections will not be missed because of genetic variation, as the platform exhibited similar limits of detection for the subtypes evaluated, meeting the sensitivity requirements set by the regulatory bodies.
authors.affiliation: Molecular Biology Centre, Department of Transfusion Medicine and Blood Bank, Serviço de Imuno-hemoterapia, Hospital S. João and Medicine Faculty of Porto University, Portugal. fernando.araujo@hsjoao.min-saude.pt
authors.forename: Fernando
authors.initials: F
authors.lastname: Araújo
authors.forename: Isabel
authors.initials: I
authors.lastname: Henriques
authors.forename: Fátima
authors.initials: F
authors.lastname: Monteiro
authors.forename: Elsa
authors.initials: E
authors.lastname: Meireles
authors.forename: Aldina
authors.initials: A
authors.lastname: Cruz
authors.forename: Gilberto
authors.initials: G
authors.lastname: Tavares
authors.forename: A
authors.initials: A
authors.lastname: Mota-Miranda
mesh_terms: D000328:Adult; D001331:Automation; D001782:Blood Donors; D001803:Blood Transfusion / Q000592:standards; D018562:Disease Transmission, Infectious / Q000517:prevention & control; D005602:France / Q000453:epidemiology; D005838:Genotype; D015658:HIV Infections / Q000097:blood* / Q000453:epidemiology / Q000635:transmission / Q000821:virology; D015497:HIV-1 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000097:blood* / Q000453:epidemiology / Q000635:transmission / Q000821:virology; D006801:Humans; D008403:Mass Screening / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D012367:RNA, Viral / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA; D012822:Silicon Dioxide; D065227:Transfusion Reaction; D014766:Viremia / Q000453:epidemiology / Q000821:virology*
chemical_list: D012367:RNA, Viral; D012822:Silicon Dioxide