DB21T 3111-2019 水質(zhì) 腸球菌的測(cè)定 固定底物技術(shù)酶底物法

DB21Waterquality—DeterminationofEnterococci—Definedsubstratetechnology遼寧省市場(chǎng)監(jiān)督管理局發(fā)布I 1 1 1 1 2 2 3 3 4 4 5 5 5 6本標(biāo)準(zhǔn)依據(jù)GB/T1.1-2009《標(biāo)準(zhǔn)化工作導(dǎo)則1水質(zhì)腸球菌的測(cè)定固定底物技術(shù)酶底物法本標(biāo)準(zhǔn)適用于地表水、地下水、飲用水、海水、凡是不注日期的引用文件，其最新版本（包括所有的修改單）GB/T5750.2生活飲用水標(biāo)準(zhǔn)檢驗(yàn)方法樣品的采集與GB/T6682分析實(shí)驗(yàn)室用水規(guī)格和GB17378.7海洋監(jiān)測(cè)規(guī)范第七部分：近海污染生態(tài)調(diào)查和HJ/T91地表水和污水監(jiān)測(cè)技術(shù)規(guī)范41℃培養(yǎng)24h，能產(chǎn)生β-半乳糖苷酶(β-D-galactosidase），分解選擇性培養(yǎng)基中的4-甲基傘形酮-β-D-葡萄糖醛酸苷（4-methyl-umbelliferyl-β-D-glucoside，MUG）釋放出熒光物質(zhì)（4-2在特定溫度下培養(yǎng)特定的時(shí)間，腸球菌能產(chǎn)生β-半乳糖苷酶，分解熒光底物釋放出熒光產(chǎn)物，在5.1培養(yǎng)基6.1恒溫培養(yǎng)箱：能夠恒溫至41℃,控溫精度0.5℃。37.2.1含活性氯（余氯）的水樣，在7.2.2含重金屬的水樣，在采樣瓶滅菌前需加入0.6ml乙二胺四根據(jù)水樣污染程度確定稀釋度，避免接種水樣經(jīng)培養(yǎng)后出現(xiàn)全部陽性或全部陰性。接種量小于100ml時(shí)，水樣應(yīng)制成稀釋樣品后使用，接種量為10ml時(shí)，以無菌操作方法取10ml水樣加入到裝有90ml無菌水的三角瓶（6.6）中混勻制成1:10（稀釋度）稀釋的樣品，其它接種量的稀釋樣品依次類推。以無菌操作方法量取100ml原水樣或稀釋），將封口后的97孔定量盤（6.7）放入培養(yǎng)箱（6.1）中培4），），定結(jié)果無效，應(yīng)重新測(cè)定。腸球菌陽性。若結(jié)果可疑，可延長培養(yǎng)時(shí)間到28h進(jìn)行結(jié)果判讀，超過28h后出現(xiàn)的熒光反應(yīng)不作為陽9結(jié)果計(jì)算與表示9.1結(jié)果計(jì)算從97孔定量盤法MPN表（附錄A）中查得每100ml水樣中腸球菌的MPN值，再根據(jù)樣品不同的稀釋度進(jìn)行計(jì)算，每100ml水樣中腸球菌的計(jì)算公式9.2結(jié)果表示測(cè)定結(jié)果保留兩位有效數(shù)字，大于等于100時(shí)以科學(xué)計(jì)數(shù)法表示，結(jié)果單位為CFU/1多家實(shí)驗(yàn)室分別對(duì)腸球菌有證標(biāo)準(zhǔn)樣品（221±37CFU/100ml）和低濃度（5CFU/100ml）、中濃度（980CFU/100ml）、高濃度(114500CFU/100ml)實(shí)際樣品進(jìn)行了測(cè)定，多家實(shí)驗(yàn)室對(duì)腸球菌有證標(biāo)準(zhǔn)樣品（221±37C-1.4%～-2.9%，相對(duì)誤差的最終值為-2.0%±1.5697孔定量盤法MPN表（CFU/100ml）大孔陽性格數(shù)小孔陽性格數(shù)<12.03.14.15.22.03.04.15.26.32.03.04.15.16.27.33.04.05.16.17.28.44.05.06.17.28.39.45.06.07.18.29.310.56.07.18.19.210.411.57.08.19.210.311.412.68.09.110.211.312.513.79.010.111.212.413.514.710.011.112.213.414.615.811.012.113.314.515.616.912.013.214.315.516.717.913.014.215.416.517.819.014.115.216.417.618.820.115.116.217.418.619.921.216.117.318.519.721.022.217.118.319.520.822.023.318.119.320.621.823.124.419.120.421.622.924.225.520.221.422.723.925.326.621.222.423.725.026.327.722.223.524.826.127.428.823.324.525.827.128.529.924.325.626.928.229.631.06.37.58.69.811.07.48.59.710.912.18.49.610.812.013.29.510.711.913.114.410.611.813.014.215.511.612.814.115.316.612.713.915.216.417.713.815.016.317.618.914.916.117.418.720.016.017.218.519.821.117.018.319.620.922.318.119.420.722.023.419.220.521.823.224.620.321.622.924.325.721.422.724.125.426.922.523.825.226.628.023.624.926.327.729.224.726.027.428.930.325.827.128.630.031.526.928.329.731.232.728.029.430.832.333.829.130.532.033.535.030.231.633.134.636.231.332.834.335.837.432.433.935.437.038.612.213.514.816.117.513.414.616.017.318.714.515.817.118.519.915.616.918.319.721.116.818.119.520.922.317.919.320.622.123.519.120.421.823.324.720.221.623.024.525.921.422.824.225.727.222.523.925.426.928.423.725.126.628.129.624.826.327.829.330.926.027.529.030.532.127.228.630.231.733.328.329.831.433.034.629.531.032.634.235.830.732.233.835.437.131.933.435.036.738.433.034.636.237.939.634.235.837.539.140.935.437.038.740.442.236.638.239.941.643.437.839.541.242.944.739.040.742.444.246.040.241.943.645.447.32018.920.321.823.324.920.121.623.124.626.221.322.824.325.927.522.624.125.627.228.823.825.326.928.530.125.026.628.129.831.526.227.829.431.132.827.529.130.732.434.128.730.332.033.735.430.031.633.335.036.831.232.934.636.338.132.534.135.937.639.533.735.437.239.040.835.036.738.540.342.236.338.039.841.643.637.539.341.143.044.938.840.642.444.346.340.141.943.845.747.741.443.245.147.149.142.744.546.548.450.544.045.947.849.851.945.347.249.251.253.346.648.550.552.654.747.949.851.954.056.149.251.253.255.457.6212223242526.528.229.931.733.627.929.531.333.135.029.230.932.734.536.430.532.334.135.937.931.833.635.537.339.333.235.036.838.840.834.536.438.340.242.235.937.739.741.743.737.339.141.143.145.238.640.542.544.646.740.041.943.946.048.241.443.345.447.549.742.844.846.849.051.244.146.248.350.552.745.547.649.752.054.346.949.051.253.555.848.450.552.755.057.349.851.954.256.558.951.253.455.658.060.552.654.857.159.562.054.156.358.661.163.655.557.860.262.665.256.959.361.764.266.858.460.863.265.868.459.962.364.767.370.0262728293035.537.439.541.743.936.938.941.043.245.538.440.442.644.847.139.942.044.146.448.741.443.545.748.050.442.845.047.349.652.044.346.548.851.253.745.948.150.452.855.447.449.652.054.557.148.951.253.656.158.850.452.855.257.860.552.054.456.959.562.253.556.058.561.264.055.157.660.262.965.756.759.261.864.667.558.260.863.566.369.359.862.465.268.071.061.464.166.969.872.963.065.768.671.574.764.767.470.373.376.566.369.172.075.178.367.970.873.776.980.269.672.575.578.782.171.274.277.380.584.072.975.979.082.485.9313233343546.248.751.253.956.847.950.453.055.758.649.552.154.857.660.551.253.856.559.462.452.955.658.361.364.454.657.360.263.166.356.359.162.065.068.358.160.963.867.070.359.862.765.768.972.361.664.567.670.874.363.366.369.572.876.365.168.271.474.878.466.970.073.376.880.568.771.975.278.882.670.573.877.280.884.772.475.779.282.986.974.277.681.285.089.176.179.583.287.191.378.081.585.289.293.579.983.587.391.495.781.885.489.393.598.083.787.591.495.7100.385.789.593.697.9102.687.691.595.7100.2105.089.693.697.8102.4107.3363738394059.862.966.370.073.861.765.068.472.276.263.767.070.674.478.565.769.172.776.780.967.771.274.978.983.369.773.377.181.385.771.775.479.483.688.273.877.681.686.090.875.979.883.988.493.378.082.086.290.995.980.184.288.693.498.582.386.591.095.9101.284.588.893.498.4103.986.791.195.8101.0106.788.993.498.3103.6109.591.295.8100.8106.3112.493.598.2103.4109.0115.395.8100.6105.9111.8118.298.1103.1108.6114.6121.2100.5105.6111.2117.4124.3102.9108.1113.9120.3127.4105.3110.7116.6123.2130.5107.7113.3119.4126.1133.7110.2115.9122.2129.2137.0112.7118.6125.0132.2140.3414243444578.082.687.693.199.380.585.290.496.1102.583.087.893.299.1105.885.590.596.0102.2109.288.093.299.0105.4112.690.696.0101.9108.6116.293.398.8105.0111.9119.895.9101.7108.1115.3123.698.7104.6111.2118.7127.4101.4107.6114.5122.3131.4104.3110.6117.8125.9135.4107.1113.7121.1129.6139.6110.0116.9124.6133.4143.9113.0120.1128.1137.4148.3116.0123.4131.7141.4152.9119.1126.7135.4145.5157.6122.2130.1139.1149.7162.4125.4133.6143.0154.1167.4128.7137.2147.0158.5172.6132.0140.8151.0163.1178.0135.4144.5155.2167.9183.5138.8148.3159.4172.7189.2142.3152.2163.8177.7195.1145.9156.1168.2182.9201.2149.5160.2172.8188.2207.546474849106.3114.3123.9135.5109.8118.3128.4140.8113.4122.4133.1146.4117.2126.6137.9152.3121.0130.9143.0158.5125.0135.4148.3165.0129.1140.1153.9172.0133.3145.0159.7179.3137.6150.0165.8187.2142.1155.3172.2195.6146.7160.7178.9204.6151.5166.4186.0214.3156.5172.3193.5224.7161.6178.5201.4235.9167.0185.0209.8248.1172.5191.8218.7261.3178.2198.9228.2275.5184.2206.4238.2290.9190.4214.2248.9307.6196.8222.4260.3325.5203.5231.0272.3344.8210.5240.0285.1365.4217.8249.5298.7387.3225.4259.5313.0410.6233.3270.0328.2435.27大孔陽性格數(shù)小孔陽性格數(shù)25262728293031323334353637383940414243444546474825.326.627.929.330.732.126.427.729.030.431.833.227.428.730.031.432.834.328.429.831.132.533.935.429.530.832.233.635.036.530.531.933.234.736.137.631.532.934.335.837.238.732.634.035.436.838.339.933.635.036.537.939.441.034.736.137.539.040.542.135.737.238.640.141.643.236.838.239.741.242.844.437.839.340.842.343.945.538.940.441.943.445.046.640.041.443.044.546.147.741.042.544.045.647.248.942.143.645.146.748.350.043.144.746.247.849.551.244.245.747.348.950.652.345.346.848.450.051.753.546.347.949.551.252.954.647.449.050.652.354.055.848.550.151.753.455.156.949.551.252.854.556.358.133.535.036.638.139.734.736.237.739.340.935.837.338.940.542.136.938.440.041.643.338.039.641.242.844.539.240.742.344.045.740.341.943.545.246.941.443.044.746.448.142.644.245.947.649.343.745.347.048.850.644.846.548.250.051.846.047.749.451.253.047.148.850.652.454.248.350.051.853.655.549.451.253.054.856.750.652.354.156.057.951.753.555.357.259.252.954.756.558.460.454.155.957.759.761.755.257.159.060.962.956.458.360.262.164.257.659.461.463.465.458.760.662.664.666.759.961.863.865.867.941.443.144.946.748.642.644.346.148.049.943.845.647.449.351.245.046.848.650.552.546.348.149.951.853.847.549.351.253.155.148.750.652.554.456.449.951.853.755.757.851.253.155.057.059.152.454.356.358.360.453.755.657.659.661.854.956.858.960.963.156.158.160.262.364.557.459.461.563.665.858.660.762.864.967.259.962.064.166.368.561.263.265.467.669.962.464.566.768.971.363.765.868.070.372.665.067.169.371.674.066.368.470.773.075.467.569.772.074.476.868.871.073.375.778.270.172.474.777.179.62050.552.554.656.859.051.853.956.058.260.453.255.257.459.661.954.556.658.861.063.355.858.060.262.464.857.259.361.663.966.358.560.763.065.367.759.962.164.466.869.261.263.565.868.270.762.664.967.269.772.264.066.368.671.173.765.367.770.172.675.266.769.171.574.176.768.170.573.075.578.269.571.974.477.079.870.973.375.978.581.372.374.877.380.082.873.776.278.881.584.475.177.680.383.185.976.579.181.884.687.577.980.583.386.189.179.382.084.887.690.780.883.586.389.292.282.284.987.890.793.8212223242561.363.866.368.971.762.865.367.870.573.364.366.869.472.175.065.868.371.073.776.667.369.872.575.378.368.871.474.177.080.070.372.975.778.681.771.874.577.380.383.373.376.178.981.985.174.977.680.583.686.876.479.282.285.288.577.980.883.886.990.279.582.485.488.692.081.184.087.190.393.782.685.688.792.095.584.287.290.493.897.385.888.992.195.599.187.490.593.897.2100.989.092.195.599.0102.790.693.897.2100.7104.592.295.598.9102.5106.393.897.1100.6104.3108.295.498.8102.4106.1110.097.1100.5104.1107.9111.9262728293074.677.680.884.287.876.379.482.686.189.778.081.184.487.991.779.782.986.389.893.681.484.688.191.795.683.186.489.993.797.684.888.291.895.699.686.690.093.797.5101.688.491.995.699.5103.790.193.797.5101.5105.791.995.599.4103.5107.893.797.4101.3105.5109.995.599.3103.3107.5112.097.3101.2105.2109.5114.299.2103.1107.2111.6116.3101.0105.0109.2113.7118.5102.9106.9111.2115.7120.6104.7108.8113.2117.8122.8106.6110.8115.2120.0125.1108.5112.7117.3122.1127.3110.4114.7119.3124.2129.5112.3116.7121.4126.4131.8114.2118.7123.5128.6134.1116.2120.7125.6130.8136.4313233343591.695.7100.0104.7109.793.697.8102.2107.0112.295.699.9104.4109.3114.697.7102.0106.6111.7117.199.7104.2108.9114.0119.6101.8106.3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