Quantifying Infectious Viruses in Water Samples Using an In Vitro Total Culturable Virus Assay

要約

Source: Fout, G. S., Cashdollar, J. L. EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay. J. Vis. Exp. (2016)

This video demonstrates the use of a total culturable virus assay to quantify infectious viruses in water samples. It outlines host cell inoculation, observation of cytopathic effect, and statistical quantification of infectious virus concentration.

プロトコル

1. Total Culturable Virus Quantal Assay
NOTE: For all steps, always add solutions carefully to avoid disturbing the cell monolayer. 

1. Decant or aspirate media from test vessels containing a monolayer of buffalo green monkey kidney (BGM) cells at 3–6 days post splitting and then add a volume of balanced salt solution equal to the media removed.
2. Decant or aspirate the balanced salt solution from the cell culture test vessels being used, and then inoculate the cell culture test vessels.
   1. Inoculate 10 test vessels for each test sample with a volume of subsample 1 equal to the Inoculum V....

材料

この記事で使用された材料の一覧

名前	会社	カタログ番号	コメント
Beef extract, desiccated powder	BD Bacto	211520
Hank’s balanced salt solution	Invitrogen	14170-112
Mechanical rocking platform	Daigger	EF4907G
Orbital shaker	Thermo Fisher	14-285-729
Sterilizing filter with prefilter	VWR	28143-295
Sterilizing syringe filter	Corning	431219
MEM	Sigma-Aldrich	M1018 or M4642
Leibovitz L-15	Sigma-Aldrich	L4386
Sodium bicarbonate, 7.5%	Sigma-Aldrich	S8761
Fetal bovine serum	Invitrogen	10082-139
Antibiotic-Antimycotic	Life Technologies	15240-062
Trypsin, EDTA	Invitrogen	25200072