期刊
JOURNAL OF VIROLOGICAL METHODS
卷 185, 期 2, 页码 267-275出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2012.07.011
关键词
HIV; Neutralizing antibody assay; Proficiency test; TZM-bl; Pseudovirus
资金
- National Science and Technology [2012ZX10004701-001]
Among the neutralizing antibody evaluation assays, the single-cycle pseudovirus infection assay is high-throughput and can provide rapid, sensitive and reproducible measurements after a single cycle of infection. Cell counts, pseudovirus inoculation levels, amount of diethylaminoethyl-dextran (DEAE-dextran), and the nonspecific effects of serum and plasma were tested to identify the optimal conditions for a neutralizing antibody assay based on pseudoviruses. Optimal conditions for cell counts, pseudovirus inoculation, and amount of DEAE-dextran were 1 x 10(4) cells/well, 200 TCID50/well, and 15 mu g/ml, respectively. Compared with serum samples, high-concentration anticoagulants reduced the relative light unit (RLU) value. The RLU value increased sharply initially but then decreased slowly with dilution of the plasma sample. Test kits containing 10 HIV-1 CRF07/08_BC pseudovirus strains and 10 plasma samples from individuals infected with HIV-1 CRF07/08_BC were assembled into two packages and distributed to nine laboratories with a standard operating procedure included. For the 10 laboratories that evaluated the test, 17 of 44 (37%) laboratory pairs were considered equivalent. A statistical qualification rule was developed based on the testing results from 5 experienced laboratories, where a laboratory qualified if at least 83% of values lied within the acceptable range. (C) 2012 Elsevier B.V. All rights reserved.
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