title: Pilot studies for development of an HIV subtype panel for surveillance of global diversity.
abstract: The continued global spread and evolution of HIV diversity pose significant challenges to diagnostics and vaccine strategies. NIAID partnered with the FDA, WRAIR, academia, and industry to form a Viral Panel Working Group to design and prepare a panel of well-characterized current and diverse HIV isolates. Plasma samples that had screened positive for HIV infection and had evidence of recently acquired infection were donated by blood centers in North and South America, Europe, and Africa. A total of 80 plasma samples were tested by quantitative nucleic acid tests, p24 antigen, EIA, and Western blot to assign a Fiebig stage indicative of approximate time from initial infection. Evaluation of viral load using FDA-cleared assays showed excellent concordance when subtype B virus was tested, but lower correlations for subtype C. Plasma samples were cocultivated with phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs) from normal donors to generate 30 viral isolates (50-80% success rate for samples with viral load >10,000 copies/ml), which were then expanded to 10(7)-10(9) virus copies per ml. Analysis of env sequences showed that sequences derived from cultured PBMCs were not distinguishable from those obtained from the original plasma. The pilot collection includes 30 isolates representing subtypes B, C, B/F, CRF04_cpx, and CRF02_AG. These studies will serve as a basis for the development of a comprehensive panel of highly characterized viral isolates that reflects the current dynamic and complex HIV epidemic, and will be made available through the External Quality Assurance Program Oversight Laboratory (EQAPOL).
authors.affiliation: SeraCare Life Sciences, Inc., Gaithersburg, Maryland, USA. mmanak@hivresearch.org
authors.forename: Mark
authors.initials: M
authors.lastname: Manak
authors.forename: Silvana
authors.initials: S
authors.lastname: Sina
authors.forename: Bharathi
authors.initials: B
authors.lastname: Anekella
authors.forename: Indira
authors.initials: I
authors.lastname: Hewlett
authors.forename: Eric
authors.initials: E
authors.lastname: Sanders-Buell
authors.forename: Viswanath
authors.initials: V
authors.lastname: Ragupathy
authors.forename: Jerome
authors.initials: J
authors.lastname: Kim
authors.forename: Marion
authors.initials: M
authors.lastname: Vermeulen
authors.forename: Susan L
authors.initials: SL
authors.lastname: Stramer
authors.forename: Ester
authors.initials: E
authors.lastname: Sabino
authors.forename: Piotr
authors.initials: P
authors.lastname: Grabarczyk
authors.forename: Nelson
authors.initials: N
authors.lastname: Michael
authors.forename: Sheila
authors.initials: S
authors.lastname: Peel
authors.forename: Patricia
authors.initials: P
authors.lastname: Garrett
authors.forename: Sodsai
authors.initials: S
authors.lastname: Tovanabutra
authors.forename: Michael P
authors.initials: MP
authors.lastname: Busch
authors.forename: Marco
authors.initials: M
authors.lastname: Schito
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D000465:Algorithms; D001938:Brazil / Q000453:epidemiology; D002478:Cells, Cultured; D016679:Genome, Viral / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology*; D006679:HIV Seropositivity / Q000453:epidemiology / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D010865:Pilot Projects; D011044:Poland / Q000453:epidemiology; D011159:Population Surveillance; D013019:South Africa / Q000453:epidemiology; D014481:United States / Q000453:epidemiology
chemical_list: D016915:AIDS Vaccines; D015483:HIV Antibodies