Table 4.
Results of the blind mixture (total mass of 150 mg) test for evaluating the reliability of the developed primer
| A | ||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Primer sets | A. capillaris and A. iwayomogi blind test | |||||||||||||||||||
| Sample number | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 |
| 18 s rRNA primer | 13.76 ± 0.146 | 13.641 ± 0.317 | 13.679 ± 0.280 | 13.878 ± 0.144 | 13.582 ± 0.190 | 13.041 ± 0.235 | 13.531 ± 0.429 | 13.225 ± 0.213 | 12.99 ± 0.000 | 13.102 ± 0.094 | 13.621 ± 0.125 | 13.507 ± 0.225 | 13.438 ± 0.285 | 13.667 ± 0.142 | 13.458 ± 0.148 | 13.618 ± 0.122 | 13.671 ± 0.115 | 13.438 ± 0.113 | 13.087 ± 0.123 | 13.099 ± 0.106 |
| A. capillaris | ||||||||||||||||||||
| ITS 26.14 cyclesa | 19.785 ± 0.109 | 21.841 ± 0.093 | 22.146 ± 0.261 | 22.216 ± 0.212 | 19.876 ± 0.109 | 21.87 ± 0.166 | 22.34 ± 0.173 | 21.277 ± 0.127 | 33.232 ± 1.478 | 20.889 ± 0.081 | 22.015 ± 0.070 | 24.702 ± 0.162 | 21.296 ± 0.052 | 32.707 ± 0.435 | 21.858 ± 0.154 | 21.055 ± 0.140 | 22.166 ± 0.191 | 23.188 ± 0.254 | 31.704 ± 0.354 | 23.459 ± 0.238 |
| accD 28.61 cycles | 22.924 ± 0.063 | 25.009 ± 0.035 | 25.859 ± 0.169 | 25.861 ± 0.111 | 23.466 ± 0.156 | 25.165 ± 0.268 | 25.72 ± 0.008 | 24.729 ± 0.240 | 34.872 ± 0.820 | 23.964 ± 0.142 | 25.521 ± 0.153 | 27.919 ± 0.068 | 24.766 ± 0.064 | 34.71 ± 0.301 | 24.795 ± 0.209 | 24.383 ± 0.096 | 25.716 ± 0.115 | 26.131 ± 0.209 | 33.891 ± 0.960 | 26.397 ± 0.085 |
| rpoB 28.35 cycles | 23.153 ± 0.091 | 25.167 ± 0.029 | 25.886 ± 0.066 | 25.931 ± 0.062 | 23.721 ± 0.026 | 25.182 ± 0.130 | 25.8 ± 0.002 | 24.919 ± 0.048 | 35.344 ND | 24.348 ± 0.185 | 25.8 ± 0.045 | 28.107 ± 0.068 | 24.832 ± 0.028 | 37.466 ± 2.853 | 24.951 ± 0.193 | 23.909 ± 0.235 | 25.879 ± 0.130 | 26.076 ± 0.160 | 33.212 ± 0.762 | 26.556 ± 0.167 |
| A. iwayomogi | ||||||||||||||||||||
| matK 30.46 cycles | 21.23 ± 0.022 | 21.144 ± 0.085 | 21.023 ± 0.022 | 21.363 ± 0.099 | 20.887 ± 0.150 | 20.731 ± 0.084 | 20.91 ± 0.051 | 20.565 ± 0.093 | 21.095 ± 0.067 | 20.558 ± 0.089 | 21.018 ± 0.032 | 20.985 ± 0.021 | 20.816 ± 0.139 | 20.985 ± 0.035 | 20.952 ± 0.046 | 20.946 ± 0.101 | 20.891 ± 0.128 | 20.812 ± 0.094 | 20.659 ± 0.021 | 20.589 ± 0.093 |
| ycf1 27.08 cycles | 23.084 ± 0.119 | 23.108 ± 0.037 | 23.334 ± 0.194 | 23.239 ± 0.177 | 23.269 ± 0.158 | 23.019 ± 0.290 | 22.979 ± 0.192 | 22.919 ± 0.079 | 23.196 ± 0.183 | 22.895 ± 0.149 | 23.137 ± 0.121 | 23.07 ± 0.111 | 23.16 ± 0.252 | 23.286 ± 0.201 | 23.137 ± 0.355 | 23.01 ± 0.147 | 22.883 ± 0.116 | 22.83 ± 0.184 | 22.793 ± 0.154 | 22.795 ± 0.193 |
| rpoB 28.51 cycles | 22.135 ± 0.105 | 22.211 ± 0.039 | 22.296 ± 0.058 | 21.942 ± 0.068 | 21.875 ± 0.131 | 21.727 ± 0.007 | 21.969 ± 0.024 | 21.841 ± 0.001 | 21.993 ± 0.012 | 21.783 ± 0.074 | 22.129 ± 0.157 | 22.026 ± 0.093 | 22.165 ± 0.052 | 22.082 ± 0.033 | 21.98 ± 0.003 | 21.877 ± 0.062 | 21.893 ± 0.112 | 21.732 ± 0.015 | 21.733 ± 0.087 | 21.704 ± 0.064 |
| Test results | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O |
| B | ||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Primer sets | A. capillaris and A. iwayomogi blind test | |||||||||||||||||||
| Sample number | 21 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | 29 | 30 | 31 | 32 | 33 | 34 | 35 | 36 | 37 | 38 | 39 | 40 |
| 18 s rRNA primer | 13.392 ± 0.091 | 13.18 ± 0.112 | 13.1 ± 0.077 | 13.094 ± 0.188 | 13.176 ± 0.096 | 12.915 ± 0.079 | 13.112 ± 0.064 | 12.904 ± 0.012 | 12.847 ± 0.115 | 13.039 ± 0.165 | 13.19 ± 0.101 | 13.37 ± 0.100 | 13.059 ± 0.140 | 13.343 ± 0.091 | 13.054 ± 0.053 | 13.22 ± 0.128 | 13.073 ± 0.205 | 13.005 ± 0.100 | 13.041 ± 0.171 | 12.81 ± 0.106 |
| A. capillaris | ||||||||||||||||||||
| ITS 26.14 cyclesa | 15.604 ± 0.377 | 14.895 ± 0.095 | 14.88 ± 0.142 | 15.596 ± 0.397 | 15.26 ± 0.049 | 14.856 ± 0.117 | 15.126 ± 0.091 | 14.883 ± 0.119 | 15.214 ± 0.094 | 14.929 ± 0.039 | 14.797 ± 0.114 | 15.582 ± 0.114 | 14.805 ± 0.093 | 15.287 ± 0.203 | 14.973 ± 0.152 | 15.194 ± 0.101 | 14.921 ± 0.146 | 15.306 ± 0.048 | 15.477 ± 0.006 | 14.98 ± 0.072 |
| accD 28.61 cycles | 18.895 ± 0.141 | 18.473 ± 0.057 | 18.402 ± 0.133 | 18.294 ± 0.081 | 18.371 ± 0.053 | 18.259 ± 0.034 | 18.287 ± 0.072 | 18.378 ± 0.037 | 18.175 ± 0.075 | 18.172 ± 0.013 | 18.206 ± 0.248 | 18.362 ± 0.111 | 18.223 ± 0.050 | 18.356 ± 0.108 | 18.188 ± 0.188 | 18.229 ± 0.035 | 18.159 ± 0.051 | 17.993 ± 0.082 | 18.106 ± 0.008 | 17.899 ± 0.039 |
| rpoB 28.35 cycles | 18.578 ± 0.050 | 17.994 ± 0.048 | 18.079 ± 0.034 | 17.997 ± 0.058 | 18.03 ± 0.027 | 18.026 ± 0.136 | 17.94 ± 0.026 | 17.955 ± 0.014 | 17.955 ± 0.009 | 17.974 ± 0.036 | 18.058 ± 0.019 | 18.26 ± 0.107 | 18.074 ± 0.030 | 18.192 ± 0.034 | 18.03 ± 0.050 | 18.09 ± 0.024 | 18.032 ± 0.052 | 17.949 ± 0.063 | 18.008 ± 0.112 | 17.888 ± 0.030 |
| A. iwayomogi | ||||||||||||||||||||
| matK 30.46 cycles | 22.863 ± 0.001 | 23.514 ± 0.061 | 24.936 ± 0.042 | 34.519 ± 1.008 | 23.772 ± 0.104 | 23.006 ± 0.151 | 26.165 ± 0.000 | 24.204 ± 0.083 | 26.98 ± 0.119 | 22.925 ± 0.011 | 22.945 ± 0.184 | 24.096 ± 0.031 | 24.448 ± 0.021 | 24.834 ± 0.085 | 26.111 ± 0.196 | 24.728 ± 0.171 | 34.742 ± 1.218 | 34.385 ± 0.581 | 24.827 ± 0.048 | 26.191 ± 0.261 |
| ycf1 27.08 cycles | 24.776 ± 0.101 | 25.059 ± 0.051 | 26.09 ± 0.190 | 34.323 ± 0.190 | 25.74 ± 0.108 | 25.174 ± 0.153 | 27.614 ± 0.198 | 25.747 ± 0.110 | 28.195 ± 0.114 | 24.513 ± 0.288 | 26.148 ± 0.176 | 25.848 ± 0.103 | 26.122 ± 0.185 | 26.075 ± 0.122 | 27.339 ± 0.183 | 26.605 ± 0.172 | 35.054 ± 0.094 | 33.2 ± 0.186 | 26.237 ± 0.150 | 28.222 ± 0.179 |
| rpoB 28.51 cycles | 24.237 ± 0.134 | 24.571 ± 0.080 | 25.817 ± 0.031 | 31.722 ± 0.020 | 24.955 ± 0.135 | 24.499 ± 0.117 | 26.966 ± 0.013 | 25.17 ± 0.194 | 27.929 ± 0.033 | 23.935 ± 0.044 | 25.701 ± 0.069 | 25.02 ± 0.062 | 25.148 ± 0.004 | 25.736 ± 0.037 | 26.795 ± 0.189 | 26.15 ± 0.006 | 32.065 ± 0.095 | 31.939 ± 0.055 | 25.68 ± 0.041 | 27.034 ± 0.009 |
| Test results | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O | O |