DB37-T 4153-2020 水质　肠球菌的测定　酶底物法

ICS13.060C51

DB37山 东 省 地 方 标 准DB37/T4153—2020水质肠球菌的测定酶底物法Waterquality—Determinationofenterococcus—Enzymesubstratemethod20202020092520201025山东省市场监督管理局发布目 次前言 II112范引文件 13法理 14剂材料 15器设备 2627析骤 28果算报告 29密和确度 310量证质控制 3附录A（范附）附录A（范附）51孔量法同性果的可数（MPN）及95%信围 4附录B（范附）97孔量法MPN表 6前 言本标准按照GB/T1.1—2009给出的规则起草。水质肠球菌的测定酶底物法范围本标准规定了测定生活饮用水及其水源水中肠球菌的酶底物法。本标准适用于生活饮用水及其水源水中肠球菌的测定。100mL1MPN/100mLGB/T5750.2—2006生活饮用水标准检验方法水样的采集与保存GB/T6682分析实验室用水规格和试验方法4124h，β-D-4-β-D366nmMPNEnterolert0.85GB/T668212120min，98（5.0g/L）：0.5g（4.5）0.1g100mL，12120min（PBS）：氯化钠 8.01氯化钾 0.2g磷酸二钠 1.44磷酸氢钾 0.24950mLpH7.41000mL，12120min100mL、500mL41℃±1121366nm51970.001g。10mL100mL样品200mL（125mL20μL）GB/T5750.2—20067分析步骤7分析步骤100mL（5.1）量取100mL（4.1），（5.2）中41±124h。100mL100mL100mL51（5.6）（5.4）（5.2）41124h选用无菌水（4.4）按照7.1～7.2步骤进行空白对照测定。8结果计算与报告定性分析（7.1）的样品，在紫外灯（5.5）下观察，产生荧光，则判定为阳性反应；不产生荧光，则判定为阴性反应。定性反应结果以肠球菌“阳性（+）”或“阴性（-）”报告。24A表A.1MPN/100mL2828h注：对于肠球菌浓度较高的样品，可选用97孔定量盘或采用无菌生理盐水（4.2）对样品1:10梯度稀释后进行多梯度检测，其他操作步骤同51孔定量盘法一致，结果检索表见附录B表B.1～表B.4，乘以稀释倍数报告。精密度6个实验室对肠球菌浓度＜30MPN/100mL的样品定性结果一致，对浓度为20.4MPN/100mL～46.3MPN/100mL、73.1MPN/100mL～146.0MPN/100m/L的样品7次重复测定，相对标准偏差分别为1.4～11.21.4～6.4准确度6166±32MPN/100mL-4.8%～0.8%范围内。注：微生物检测数据为偏态分布，上述精密度、准确度均为对测定结果以10取对数后进行计算。7.310MPN/100mL7.1～7.2对每批次培养基须选用有证标准菌株或标准样品进行培养基质量检验。11废物处理实验中产生的废弃物经高压蒸汽灭菌器（5.3）121℃灭菌20min后，作为一般废物处理。附录A（规范性附录）51孔定量盘法不同阳性结果的最可能数（MPN）及95%可信范围51孔定量盘法不同阳性结果的最可能数（MPN）及95%可信范围见表A.1。表A.151（MPN）95阳性数肠球菌MPN/100mL95%置信范围MPN/100mL下限上限0＜10.03.711.00.35.622.00.67.333.11.19.044.21.710.755.32.312.366.43.013.977.53.715.588.74.517.199.95.318.81011.16.120.51112.47.022.11213.77.923.91315.08.825.71416.49.827.51517.810.829.41619.211.931.31720.713.033.31822.214.135.21923.815.337.32025.416.539.42127.117.741.62228.819.043.92330.620.446.32432.421.848.72534.423.351.22636.424.753.92738.426.456.62840.628.059.5A.151（MPN）95阳性数肠球菌MPN/100mL95%置信范围MPN/100mL下限上限2942.929.762.53045.331.565.63147.833.469.03250.435.472.53353.137.576.23456.039.780.13559.142.084.43662.444.688.83765.947.293.73869.750.099.03973.853.1104.84078.256.4111.24183.159.9118.34288.563.9126.24394.568.2135.444101.373.1146.045109.178.6158.746118.485.0174.547129.892.7195.048144.5102.3224.149165.2115.2272.250200.5135.8387.651＞200.5146.1—附录B（）97MPN97孔定量盘法MPN表见表B.1～表B.4。表B.197PN表（N/100L011）大孔阳性数小孔阳性数012345678910110＜1.01.02.03.04.05.06.07.08.09.010.011.011.02.03.04.05.06.07.18.19.110.111.112.122.03.04.15.16.17.18.19.210.211.212.213.333.14.15.16.17.28.29.210.311.312.413.414.544.15.26.27.28.39.310.411.412.513.514.615.655.26.37.38.49.410.511.512.613.714.715.816.966.37.48.49.510.611.612.713.814.916.017.018.177.58.59.610.711.812.813.915.016.117.218.319.488.69.710.811.913.014.115.216.317.418.519.620.799.810.912.013.114.215.316.417.618.719.820.922.01011.012.113.214.415.516.617.718.920.021.122.323.41112.213.414.515.616.817.919.120.221.422.523.724.81213.514.615.816.918.119.320.421.622.823.925.126.31314.816.017.118.319.520.621.823.024.225.426.627.81416.117.318.519.720.922.123.324.525.726.928.129.31517.518.719.921.122.323.524.725.927.228.429.630.91618.920.121.322.623.825.26.227.528.730.031.232.51720.321.622.824.125.326.627.829.130.331.632.934.11821.823.124.325.626.928.129.430.732.033.334.635.91923.324.625.927.228.529.831.132.433.735.036.337.62024.926.227.528.830.131.532.834.135.436.838.139.52126.527.929.230.531.833.234.535.937.338.640.041.42228.229.530.932.333.635.036.437.739.140.541.943.32329.931.332.734.135.536.838.339.741.142.543.945.42431.733.134.535.937.338.840.241.743.144.646.047.52533.635.036.437.939.340.842.243.745.246.748.249.72635.536.938.439.941.442.844.345.947.448.950.452.02737.438.940.442.043.545.046.548.149.651.252.854.42839.541.042.644.145.747.348.850.452.053.655.256.9表B.197N表MPN100L小孔～11（）大孔阳性数小孔阳性数012345678910112941.743.244.846.448.049.651.252.854.556.157.859.53043.945.547.148.750.452.053.755.457.158.860.562.23146.247.949.551.252.954.656.358.159.861.663.365.13248.750.452.153.855.657.359.160.962.764.566.368.23351.253.054.856.558.360.262.063.865.767.669.571.43453.955.757.659.461.363.165.067.068.970.872.874.83556.858.660.562.464.466.368.370.372.374.376.378.43659.861.763.765.767.769.771.773.875.978.080.182.33762.965.067.069.171.273.375.477.679.882.084.286.53866.368.470.672.774.977.179.481.683.986.288.691.03970.072.274.476.778.981.383.686.088.490.993.495.94073.876.278.580.983.385.788.290.893.395.998.5101.24178.080.583.085.588.090.693.395.998.71001.4104.3107.14282.685.287.890.593.296.098.8101.7104.6107.6110.6113.74387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.14493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.64599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.646106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.547114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.448123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.049135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3表B.2表B.297PN表（N/100L1～2）大孔阳性数小孔阳性数121314151617181920212223012.013.014.115.116.117.118.119.120..221.222.224.3113.214.215.216.217.318.319.320.421.422.423.524.5214.315.416.417.418.519.520.621.622.723.724.825.8315.516.517.618.619.720.821.822.923.925.026.127.1416.717.818.819.921.022.023.124.225.326.327.428.5517.919.020.121.222.223.324.425.526.627.728.829.9619.220.321.422.523.624.725.826.928.029.130.231.3720.521.622.723.824.926.027.128.329.430.531.632.8821.822.924.125.226.327.428.629.730.832.033.134.3923.224.325.426.627.728.930.031.232.333.534.635.81024.625.726.928.029.230.331.532.733.835.036.237.41126.027.228.329.530.731.933.034.235.436.637.839.0表B.297N表MPN100L小孔2～23（）大孔阳性数小孔阳性数1213141516171819202122231227.528.629.831.032.233.434.635.837.038.239.540.71329.030.231.432.633.835.036.237.538.739.941.242.41430.531.733.034.235.436.737.939.140.441.642.944.21532.133.334.635.837.138.439.640.942.243.444.746.01633.735.036.337.538.840.141.442.744.045.346.647.91735.436.738.039.340.641.943.244.545.947.248.549.81837.238.539.841.142.443.845.146.547.849.250.551.91939.040.341.643.044.345.747.148.449.851.252.654.02040.842.243.644.946.347.749.150.551.953.354.756.12142.844.145.546.948.449.851.252.654.155.556.958.42244.846.247.649.050.551.953.454.856.357.859.360.82346.848.349.751.252.754.255.657.158.660.261.763.22449.050.552.053.555.056.558.059.561.162.664.265.82551.252.754.355.857.358.960.562.063.665.266.868.42653.555.156.758.259.861.463.064.766..367.969.671.22756.057.659.260.862.464.165.767.469.170.872.574.22858.560.261.863.565.266.968.670.372.073.775.577.32961.262.964.666.368.069.871.573.375.176.978.780.53064.065.767.569.371.072.974.776.578.380.282.184.03166.968.770.572.474.276.178.079.981.883.785.787.63270.071.973.875.777.679.581.583.585.487.589.591.53373.375.277.279.281.283.285.287.389.391.493.695.73476.878.880.882.985.087.189.291.493.595.797.9100.23580.582.684.786.989.191.393.595.798.0100.3102.6105.03684.586.788.991.293.595.898.1100.5102.9105.3107.7110.23788.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.93893.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.23998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.240103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.041110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.942116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.143124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.244133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.945143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.246156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.447172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.548193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.049224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6表B.397PN表（N/100L2～3）大孔阳性数小孔阳性数242526272829303132333435024.325.326.427.428.429.530.531.532.633.634.735.7125.626.627.728.729.830.831.932.934.035.036.137.2226.927.929.030.031.132.233.234.335.436.537.538.6328.229.330.431.432.533.634.735.836.839.739.040.1429.630.731.832.833.935.036.137.238.339.440.541.6531.032.133.234.335.436.537.638.739.941.042.143.2632.433.534.735.836.938.039.240.341.442.643.744.8733.935.036.237.338.439.640.741.943.044.245.346.5835.436.637.738.940.041.242.343.544.745.947.048.2937.038.139.340.541.642.844.045.246.447.648.850.01038.639.740.942.143.344.545.746.948.149.350.651.81140.241.442.643.845.046.347.548.749.951.252.453.71241.943.144.345.646.848.149.350.651.853.154.355.61343.644.946.147.448.649.951.252.553.755.056.357.61445.446.748.049.350.551.853.154.455.757.058.359.61547.348.649.951.252.553.855.156.457.859.160.461.81649.250.551.853.254.555.857.258.559.961.232.664.01751.252.553.955.256.658.059.360.762.163.564.966.31853.264.656.057.458.860.261.663.064.465.867.268.61955.456.858.259.661.062.463.965.366.868.269.771.12057.659.060.461.963.364.866.367.769.270.772.273.72159.961.332.834.365.867.368.870.371.873.374.976.42262.363.865.366.868.369.871.472.974.576.177.679.22364.766.367.869.471.072.574.175.777.378.980.582.22467.368.970.572.173.775.377.078.680.381.983.685.22570.071.773.375.076.678.380.081.783.385.186.888.52672.974.676.378.079.781.483.184.886.688.490.191.92775.977.679.481.182.984.686.488.290.091.993.795.52879.080.882.684.486.388.189.991.893.795.697.599.42982.484.286.187.989.891.793.795.697.599.5101.5103.53085.987.889.791.793.695.697.699.6101.6103.7105.7107.83189.691.693.695.697.799.7101.8103.9106.0108.2110.3112.53293.695.797.899.9102.0104.2106.3108.5110.7113.0115.2117.53397.8100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.934102.4104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.835107.3109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3表B.397N表MPN100L小孔4～35（）大孔阳性数小孔阳性数24252627282930313233343536112.7115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.437118.6121.324.0126.8129.6132.4135.3138.2141.2144.2147.3150.338125.0127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.239132.2135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.440140.3143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.141149.5153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.842160.2164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.443172.8177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.844188.2193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.145207.5214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.346233.3241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.347270.0280.9292.4304.4316.9330.0343.6357.8372.5387.740.4419.848328.2344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.349435.2461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4表B.497PN表（N/100L3～4）大孔阳性数小孔阳性数36373839404142434445464748036.837.838.940.041.042.143.144.245.346.347.448.549.5138.239.340.441.442.543.544.745.746.847.949.050.151.2239.740.841.943.044.045.146.247.348.449.550.651.752.8341.242.343.444.545.646.747.848.950.051.252.353.454.5442.843.945.046.147.248.349.550.651.752.954.055.156.3544.445.546.647.748.950.051.252.353.554.655.856.958.1646.047.148.349.450.651.752.954.155.256.457.658.759.9747.748.850.051.252.353.554.755.957.158.359.460.661.8849.450.651.853.054.155.356.557.759.060.261.462.663.8951.252.453.654.856.057.258.459.760..962.163.464.665.81053.054.255.556.757.959.260.461.762.964.265.466.767.91154.956.157.458.659.961.262.463.765.066.367.568.870.11256.858.159.460.762.063.264.565.867.168.169.771.072.41358.960.261.562.864.165.466.768.069.370.772.073.374.71460.962.363.664.966.367.668.970.371.673.074.475.777.11563.164.565.867.268.569.971.372.674.075.476.878.279.61665.366.768.169.570.972.373.775.176.577.979.380.882.2表B.497N表MPN100L小孔6～48（）大孔阳性数小孔阳性数363738394041424344454647481767.769.170.571.973.374.876.277.679.180.582.083.384.91870.171.573.074.475.977.378.880.381.883.384.886.387.81972.674.175.577.078.580.081.583.184.686.187.689.290.72075.276.778.279.881.382.884.485.987.589.190.792.293.82177.979.581.182.684.285.887.489.090.692.293.895.497.12280.882.484.085.687.288.990.592.193.895.597.198.8100.52383.885.487.188.790.492.193.895.597.298.9100.6102.4104.12486.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92590.292.093.795.5/r/