Infectious Disease Research
Virology
Retroviruses
Confirmatory Anti-HIV Evaluation
Prior to XTT staining of the microtiter plates, a cell-free supernatant sample is collected. This sample is used to quantitate virus growth in each well of the assay plate. These data (measurement of virus growth) supplement and confirm the data obtained in the primary anti-HIV assay (measurement of inhibition of virus-induced cell killing).
Assays performed include the following:
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Reverse Transcriptase Assay:
Virus content is measured using a homopolymer poly rA:oligo dT template primer system by quantitation of incorporation of tritiated thymidine monophosphate. Assays are performed in a 96-well format and quantitated using a Matrix 96 scintillation counter. A high-throughput robotic assay has been developed and implemented as well.
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p24 ELISA:
Plates used for quantitation of core protein are purchased commercially from Coulter Retrovirology. Quantitation of p24 concentration is performed using SoftMax software. A high-throughput robotic assay has been developed and implemented as well.
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Infectious virus:
Virus infectivity is measured using the CEM-SS plaque formation assay, HeLa-CD4-LTR-ß-galactosidase assays or acute infection of PBMCs. Infectious virus is measured by a variety of methods including colorimetric, microscopically and by chemiluminescence.
Anti-HIV Evaluation
Confirmatory HIV Assays
Secondary Biological Assays
Topical Microbicide Program
Combination Assays
