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Veterinary testing and diagnosis field
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Home News Veterinary Testing and Diagnosis Field

Veterinary Testing and Diagnosis Field

2024-04-23 05:47:31

Instructions for loading FMD LB-ELISA (new mode testing 10 samples).

1. As shown in the figure, A1-A10 plus 175ul of washing solution (1×PBST), A11, A12, B12 plus 100ul of washing solution (1×PBST), B1-B11, C1-C11, D1-D11, E1-E11, F1-F11, G1-G11, H1-H11 plus 100ul of washing solution (1×PBST).

  1 2 3 4 5 6 7 8 9 10 11 12
A 175ul 175ul 175ul 175ul 175ul 175ul 175ul 175ul 175ul 175ul 100ul 100ul
B 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul
C 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  
D 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  
E 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  
F 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  
G 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  
H 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul 100ul  

2. A1-A10 plus 25ul of serum to be tested, A11 plus 100ul of positive control, A12 plus 100ul of negative control, piped and mixed with a row of guns (6-7 times per well), 1-10 columns of serum to be tested were continuously diluted from A to H with an amount of 100ul, positive control 1 column was continuously diluted from A to H with an amount of 100ul, and 2 columns of negative control were diluted from A to B with an amount of 100ul.

  1 2 3 4 5 6 7 8 9 10 11 12
A 25ul 25ul 25ul 25ul 25ul 25ul 25ul 25ul 25ul 25ul 100ul 100ul
B                        
C                        
D                        
E                        
F                        
G                        
H                        

3. The dilution ratio of each well is as shown in the figure:

  1 2 3 4 5 6 7 8 9 10 11 12
A 1:08 1:08 1:08 1:08 1:08 1:08 1:08 1:08 1:08 1:08 1:16 1:02
B 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:32 1:04
C 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 0.08611  
D 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.13056  
E 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.21944  
F 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.39722  
G 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.75278  
H 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 1.46389  

4.Transfer the plate from the serum dilution plate to the corresponding well of the coated plate at 50 ul per well. Add the diluted viral antigen, add 50ul to each column 1-11, 50ul to each of A12 and B12, and 100ul to E12, F12, G12 and H12.

  1 2 3 4 5 6 7 8 9 10 11 12
A 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul
B 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul
C 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul  
D 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul  
E 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 100ul
F 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 100ul
G 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 100ul
H 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 50ul 100ul

5.After the sample is completed, the amount of each well is 100ul, and the dilution of the serum, positive control, and negative control to be tested is doubled, as shown in the figure:

  1 2 3 4 5 6 7 8 9 10 11 12
A 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:16 1:32 1:04
B 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 1:32 0.08611 1:08
C 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.08611 0.13056  
D 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.13056 0.21944  
E 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.21944 0.39722  
F 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.39722 0.75278  
G 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 0.75278 1.46389  
H 1.46389 1.46389 1.46389 1.46389 1.46389 1.46389 1.46389 1.46389 1.46389 1.46389 2.88611  
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