RESUMO
The plaque assay has long served as the "gold standard" to measure virus infectivity and test antiviral drugs, but the assay is labor-intensive, lacks sensitivity, uses excessive reagents, and is hard to automate. Recent modification of the assay to exploit flow-enhanced virus spread with quantitative imaging has increased its sensitivity. Here we performed flow-enhanced infection assays in microscale channels, employing passive fluid pumping to inoculate cell monolayers with virus and drive infection spread. Our test of an antiviral drug (5-fluorouracil) against vesicular stomatitis virus infections of BHK cell monolayers yielded a two-fold improvement in sensitivity, relative to the standard assay based on plaque counting. The reduction in scale, simplified fluid handling, image-based quantification, and higher assay sensitivity will enable infection measurements for high-throughput drug screening, sero-conversion testing, and patient-specific diagnosis of viral infections.
Assuntos
Técnicas Analíticas Microfluídicas/instrumentação , Infecções por Rhabdoviridae/tratamento farmacológico , Vírus da Estomatite Vesicular Indiana/crescimento & desenvolvimento , Vírus da Estomatite Vesicular Indiana/imunologia , Ensaio de Placa Viral/normas , Animais , Antivirais/farmacologia , Técnicas de Cultura de Células , Linhagem Celular , Cricetinae , Relação Dose-Resposta a Droga , Desenho de Equipamento , Corantes Fluorescentes/metabolismo , Fluoruracila/farmacologia , Imuno-Histoquímica , Indóis/metabolismo , Técnicas Analíticas Microfluídicas/métodos , Microtecnologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/virologia , Sensibilidade e Especificidade , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacosRESUMO
A hand-held corona treater is presented as an effective, inexpensive, and portable alternative for irreversible bonding of polydimethylsiloxane (PDMS) to glass.
