重组C因子法检测猪源纤维蛋白贴DBT细菌内毒素

Establishment of Recombinant Factor C Method for Detection of Bacterial Endotoxin in DBT

目的采用重组C因子法建立用于定量检测猪源纤维蛋白贴DBT的检测细菌内毒素方法。方法分别采用凝胶法和重组C因子法对猪源纤维蛋白贴DBT进行细菌内毒素检查。结果猪源纤维蛋白贴DBT稀释8倍后,使用Lonza和Hyglos的细菌内毒素重组C因子检测试剂盒进行干扰试验,回收率均在50%~200%之间,符合中国药典2020年版9251的规定,其限值为<3 EU·mg^(-1)。采用细菌内毒素重组C因子试剂盒按上述标准对10批猪源纤维蛋白贴DBT进行细菌内毒素检查,结果与细菌内毒素凝胶法检测结果相一致。重组C因子更有利于排除干扰,且更方便进行细菌内毒素的定量测定。结论重组C因子法能有效检测猪源纤维蛋白贴DBT的细菌内毒素,有望成为凝胶法检测的替代方法。

OBJECTIVE To establish a bacterial endotoxin detection method for quantitative detection of porcine fibrin sealant patch(DBT) using recombinant C factor method.METHODS The bacterial endotoxin test of porcine fibrin sealant patch(DBT) was carried out by gel method and recombinant factor C method respectively.RESULTS After DBT test solution was diluted 8 times,the bacterial endotoxin recombinant factor C detection kit of Lonza and Hyglos were used for interference test.The recovery rates were between 50% and 200 %,which were in line with the provisions of Chinese Pharmacopoeia(2020) 9251 guideline.The endotoxin limit was less than 3 EU·mg~(-1).The bacterial endotoxin test of 10 batches of DBT was carried out by recombinant factor C kit according to the above standards,and the results were consistent with bacterial endotoxin gel method.Recombinant factor C was more conducive to eliminating interference and more convenient for the quantitative determination of bacterial endotoxin.CONCLUSION Recombinant factor C method can effectively detect the bacterial endotoxin of DBT,which is expected to be an alternative method of gel method.