Lecture _6 - Quantifying Viral Infection & Viral...

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Quantifying Viral Infection READING: Acheson Chapter1 SARS article (web) CCD article (web) HOMEWORK (web)
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Virus concentration in virions per milliliter = 10 7 x HA titer .
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Virus Quantifcation InFectivity : quantitative versus quantal assays Quantitative Assays: (e.g. plaque assay) • Each inFectious particle can be assayed directly as a “plaque- Forming unit” (P±U) • Utilize the principle that each viable virus multiplies to produce a localized area oF cells destroyed or killed (plaque) A. Pseudorabies virus plaque (leFt) with lacZ reporter (right) on BHK cells B. Two strains oF in²uenza on the same cell type C. Poliovirus plaques on Hela cells
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Plaque Assays 2. Plate serial dilutions on susceptible monolayer of cells on a plate, allow to attach. 1. Make a 1:10 dilution series of the viral innoculum (eg, 10 , 10 , 10 . ...) -1 -2 -3 3. Overlay the cells with agar to prevent diffusion of virus particles. 4. Allow infection to proceed, resulting in localized killing of cells to form plaques; count the plaques at each dilution. Calculation of PFU/ml PFU/ml = # plaques x 10 x dilution factor plate 0.1 mL Titer = 10 plaques x10 x10 =10 PFU/mL 6 8
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What is the titer? Calculation of PFU/ml PFU/ml = # plaques x 10 x dilution factor
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Quantal Assays: (e.g. ID 50 , LD 50 , TCID 50 ) • Infectivity assays; measure the amount of viral inoculum required for a certain
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Lecture _6 - Quantifying Viral Infection & Viral...

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