Basic HTML Version
Molecular Pathogens
MP2011, Vol.2, No.3
http://mp.sophiapublisher.com
- 20 -
BP210 antisera were determined by indirect ELISA.
Serial dilutions of the antisera up to 1:16,000 were
reacted with dead and live cells of
Xac
BP210 at
10
4
~10
6
CFU/mL. The suspension of
E.coli
at
10
5
CFU/mL and pre-immune sera were used as
negative control. The results showed that, 1:4,000 or
lower dilutions of both antisera were able to detected
dead cells of
Xac
at 10
5
CFU/mL or more. Whereas,
1:2,000 diluted antiserum were reactive against live
cells of 10
6
CFU/mL and dead cells at 10
5
CFU/mL or
more. The highly concentrate antiserum (1:1,000
dilution) strongly reacted against both live and dead
cells (10
5
CFU/mL) of
Xac
. However, antiserum 1
showed stronger reaction than antiserum 2 (Table 1).
These results indicated that, at least 1:2,000 dilution
of these antisera were able to detected dead cell of
Xac
at 10
5
CFU/mL and live cells at 10
6
CFU/mL.
Nevertheless, dead cells of bacteria showed stronger
reaction with antisera than live cells. Since these
antisera were produced from injection with heat killed
cells which might have broken some cells.
Table 1 ELISA for antiserum 1 and antiserum 2 titer test
Note: Different antisera dilutions were used to detect live and dead cells of different concentrations
of
Xac
suspension; Pre-immune
serum reaction and
E. coli
at 10
5
CFU/mL was used as negative control; 100 µL of the cells were used in each well;
a
: The positive
result (+) was indicated when A
405
was greater than twice of the negative control;
b
: The negative reaction (-) was indicated when A
405
was lower than twice of the negative control
Antiserum 1
Antiserum 2
Antiserum dilution
Cell status
Xac
(CFU/mL)
A
405
Reaction
A
405
Reaction
10
6
0.25
+
a
0.20
+
10
5
0.20
+
0.18
+
Live
10
4
0.01
-
b
0.12
-
10
6
0.74
+
0.69
+
10
5
0.24
+
0.22
+
1:1,000
Dead
10
4
0.12
-
0.14
-
10
6
0.20
+
0.18
+
10
5
0.15
-
0.12
-
Live
10
4
0.10
-
0.11
-
10
6
0.75
+
0.64
+
10
5
0.21
+
0.18
+
1:2,000
Dead
10
4
0.11
-
0.12
-
Live
10
6
0.16
-
0.12
-
10
5
0.12
-
0.11
-
10
4
0.09
-
0.11
-
Dead
10
6
0.67
+
0.61
+
10
5
0.19
+
0.16
-
1:4,000
10
4
0.11
-
0.11
-
Live
10
6
0.12
-
0.10
-
10
5
0.11
-
0.10
-
10
4
0.09
-
0.10
-
Dead
10
6
0.53
+
0.51
+
10
5
0.17
-
0.14
-
1:8,000
10
4
0.10
-
0.10
-
Live
10
6
0.10
-
0.09
-
10
5
0.10
-
0.09
-
10
4
0.09
-
0.09
-
Dead
10
6
0.43
+
0.43
+
10
5
0.16
-
0.13
-
1:16,000
10
4
0.11
-
0.09
-