title: Development and implementation of an international proficiency testing program for a neutralizing antibody assay for HIV-1 in TZM-bl cells.
abstract: Recent advances in assay technology have led to major improvements in how HIV-1 neutralizing antibodies are measured. A luciferase reporter gene assay performed in TZM-bl (JC53bl-13) cells has been optimized and validated. Because this assay has been adopted by multiple laboratories worldwide, an external proficiency testing program was developed to ensure data equivalency across laboratories performing this neutralizing antibody assay for HIV/AIDS vaccine clinical trials. The program was optimized by conducting three independent rounds of testing, with an increased level of stringency from the first to third round. Results from the participating domestic and international laboratories improved each round as factors that contributed to inter-assay variability were identified and minimized. Key contributors to increased agreement were experience among laboratories and standardization of reagents. A statistical qualification rule was developed using a simulation procedure based on the three optimization rounds of testing, where a laboratory qualifies if at least 25 of the 30 ID50 values lie within the acceptance ranges. This ensures no more than a 20% risk that a participating laboratory fails to qualify when it should, as defined by the simulation procedure. Five experienced reference laboratories were identified and tested a series of standardized reagents to derive the acceptance ranges for pass-fail criteria. This Standardized Proficiency Testing Program is the first available for the evaluation and documentation of assay equivalency for laboratories performing HIV-1 neutralizing antibody assays and may provide guidance for the development of future proficiency testing programs for other assay platforms.
authors.affiliation: Duke University Medical Center, Durham, NC 27705, USA. chris.todd@duke.edu
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Todd
authors.forename: Kelli M
authors.initials: KM
authors.lastname: Greene
authors.forename: Xuesong
authors.initials: X
authors.lastname: Yu
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Ozaki
authors.forename: Hongmei
authors.initials: H
authors.lastname: Gao
authors.forename: Yunda
authors.initials: Y
authors.lastname: Huang
authors.forename: Maggie
authors.initials: M
authors.lastname: Wang
authors.forename: Gary
authors.initials: G
authors.lastname: Li
authors.forename: Ronald
authors.initials: R
authors.lastname: Brown
authors.forename: Blake
authors.initials: B
authors.lastname: Wood
authors.forename: M Patricia
authors.initials: MP
authors.lastname: D'Souza
authors.forename: Peter
authors.initials: P
authors.lastname: Gilbert
authors.forename: David C
authors.initials: DC
authors.lastname: Montefiori
authors.forename: Marcella
authors.initials: M
authors.lastname: Sarzotti-Kelsoe
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000032:analysis* / Q000276:immunology; D045744:Cell Line, Tumor; D015483:HIV Antibodies / Q000032:analysis* / Q000276:immunology; D015658:HIV Infections / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D059021:Laboratory Proficiency Testing / Q000592:standards*; D009500:Neutralization Tests / Q000592:standards*; D011786:Quality Control; D012015:Reference Standards
chemical_list: D016915:AIDS Vaccines; D057134:Antibodies, Neutralizing; D015483:HIV Antibodies