TABLE 3.
Effect of tlh primer limiting on the detection of pathogenic V. parahaemolyticus bacteria in low ratio to nonpathogenic V. parahaemolyticus bacteria
| Final concn of tlh primer and CFU/reaction of target gene: | Ratio of tdh/trh to tlh |
CT ± SDa for:
|
||||
|---|---|---|---|---|---|---|
| tdh/trh | tlh | tdh | trh | tlh | IAC | |
| 75 nM | ||||||
| 4,400 | 70,000 | 1:15 | 23.62 ± 0.33 | 24.41 ± 0.87 | 20.76 ± 0.47 | 20.31 ± 0.21 |
| 440 | 70,000 | 1:150 | 29.44 ± 0.17 | 30.83 ± 0.43 | 21.01 ± 0.14 | 20.34 ± 0.43 |
| 44 | 70,000 | 1:1,500 | 33.31 ± 0.98 | 33.82 ± 0.13 | 21.19 ± 0.08 | 20.41 ± 0.23 |
| 4.4 | 70,000 | 1:15,000 | 36.36 ± 0.49 | 37.47 ± 1.44b | 21.06 ± 0.22 | 20.18 ± 0.05 |
| 0.44 | 70,000 | 1:150,000 | 0.00 ± 0.00 | 0.00 ± 0.00 | 21.19 ± 0.02 | 19.97 ± 0.24 |
| 200 nM | ||||||
| 4,400 | 70,000 | 1:15 | 24.31 ± 1.79 | 24.40 ± 0.88 | 21.13 ± 0.08 | 20.29 ± 0.17 |
| 440 | 70,000 | 1:150 | 40.40 ± 0.77b | 0.00 ± 0.00 | 21.14 ± 0.07 | 20.37 ± 0.23 |
| 44 | 70,000 | 1:1,500 | 0.00 ± 0.00 | 0.00 ± 0.00 | 21.23 ± 0.11 | 20.23 ± 0.10 |
| 4.4 | 70,000 | 1:15,000 | 0.00 ± 0.00 | 0.00 ± 0.00 | 21.23 ± 0.02 | 20.02 ± 0.22 |
| 0.44 | 70,000 | 1:150,000 | 0.00 ± 0.00 | 0.00 ± 0.00 | 21.16 ± 0.08 | 19.90 ± 0.08 |
a
SD is the standard deviation of the CT values based on the results for three replicates.
b
Only two replicates generated a CT value; the SD was calculated excluding the negative replicate.