Author: TERAMICHI, Takurou; FUKUSHI, Shuetsu; HACHIYA, Yuma; MELAKU, Simenew Keskes; OGUMA, Keisuke; SENTSUI, Hiroshi
                    Title: Evaluation of serological assays available in a biosafety level 2 laboratory and their application for survey of Middle East respiratory syndrome coronavirus among livestock in Ethiopia  Document date: 2019_11_5
                    ID: vijh6x1l_8
                    
                    Snippet: The neutralization test using VSV-MERS/GFP was performed as previously described [8] . A medium composed of Eagle's MEM supplemented with 5% FBS was used for virus and serum dilution. Serially diluted 0.05 ml of test sera were mixed with equal volumes of 3,000 FFU of VSV-MERS/GFP and incubated at 37°C for 1 hr. The mixture was inoculated to Vero cells seeded on a 96-well culture plate and incubated at 37°C for 24 hr in a CO 2 incubator. GFP-pos.....
                    
                    
                    
                     
                    
                    
                    
                    
                        
                            
                                Document: The neutralization test using VSV-MERS/GFP was performed as previously described [8] . A medium composed of Eagle's MEM supplemented with 5% FBS was used for virus and serum dilution. Serially diluted 0.05 ml of test sera were mixed with equal volumes of 3,000 FFU of VSV-MERS/GFP and incubated at 37°C for 1 hr. The mixture was inoculated to Vero cells seeded on a 96-well culture plate and incubated at 37°C for 24 hr in a CO 2 incubator. GFP-positive cells were then detected using a fluorescence microscope. The number of positive GFP cells was counted as described above. The neutralization titer was determined as the highest serum dilution showing ≤50% of the number of GFP-positive cells compared with the no serum control.
 
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