Kathleen M Hall

US Patent:

6218105, Apr 17, 2001

Filed:

Jan 7, 2000

Appl. No.:

9/479825

Inventors:

Kathleen S. Hall - Stateline NV
Lloyd H. Smith - Davis CA

International Classification:

C12Q 168
C12Q 170
C12P 1934

US Classification:

435 5

Abstract:

A high throughput virus in vitro infectivity assay method comprising growing cells in a multi-well format, infecting the cells with intact virion incubated with a test agent, and measuring expression of at least one viral nucleic acid sequence in the cells. The method also preferably comprises incubating intact virion without test agent to define a control. The cells are preferably human keratinocyte cells grown in monolayers. The viral nucleic acid sequence will generally comprise viral mRNA. In one preferred embodiment, the intact virion comprise Human Papilloma Virus, and more preferably Human Papilloma Virus-11. Measuring expression is generally carried out by releasing the viral mRNA from the cells by lysis, amplifying the mRNA as CDNA via RT-PCR, and detecting amplicons with specific probes. Cell lysis may be carried out by heating or by treatment with detergent.