title: Detection of HIV-1 in alternative specimen types using the APTIMA HIV-1 RNA Qualitative Assay.
abstract: Peripheral blood mononuclear cells (PBMCs), saliva, seminal plasma, and dried blood spots were evaluated as specimen types for the APTIMA HIV-1 RNA Qualitative Assay (APTIMA HIV-1 Assay), which employs a target capture step to recover HIV-1-specific sequences from complex specimen types. Analytical sensitivity studies were carried out using samples that were either diluted or eluted with a buffered detergent and spiked with different concentrations of HIV-1 ranging from 1 to 10,000 copies/mL. PBMC samples spiked with HIV-1 had comparable analytical sensitivity to HIV-1 spiked plasma with a 95% limit of detection of 13.1 and 17.2 copies/mL, respectively. Analytical sensitivity in seminal plasma specimens diluted 1:5 and saliva diluted 1:2 was comparable to HIV-1 spiked dilution buffer alone. Whole blood and dried blood spot specimens spiked with HIV-1 had equivalent reactivity at 250 copies/spot (5000 copies/mL). However, the 95% limit of detection values were significantly different (293.7 copies/mL for whole blood and 2384 copies/mL for dried blood spot specimens). No significant effect on analytical sensitivity was observed when one HIV-1 positive dried blood spot punch was pooled with up to 9 HIV-1 negative dried blood spot punches. Together, these studies demonstrate that the APTIMA HIV-1 RNA Qualitative Assay can be used to process a diverse array of specimen types with minimal impact on analytical sensitivity for most specimen types.
authors.affiliation: Gen-Probe Incorporated, San Diego, CA, USA. tomn@gen-probe.com
authors.forename: C Thomas
authors.initials: CT
authors.lastname: Nugent
authors.forename: J
authors.initials: J
authors.lastname: Dockter
authors.forename: F
authors.initials: F
authors.lastname: Bernardin
authors.forename: R
authors.initials: R
authors.lastname: Hecht
authors.forename: D
authors.initials: D
authors.lastname: Smith
authors.forename: E
authors.initials: E
authors.lastname: Delwart
authors.forename: C
authors.initials: C
authors.lastname: Pilcher
authors.forename: D
authors.initials: D
authors.lastname: Richman
authors.forename: M
authors.initials: M
authors.lastname: Busch
authors.forename: C
authors.initials: C
authors.lastname: Giachetti
mesh_terms: D001801:Blood Stains; D015658:HIV Infections / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D012463:Saliva / Q000821:virology; D012661:Semen / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic