目的:建立β-内酰胺类抗生素微生物限度检查方法。方法:采用薄膜过滤并在培养基中加入β-内酰胺酶的方法,去除β-内酰胺类抗生素的抗菌活性。结果:本方法满足中国药典2005年版验证试验的基本要求。5株验证菌株中枯草芽孢杆菌对β-内酰胺类抗生素最敏感,可作为β-内酰胺类抗生素微生物限度检查方法的质控菌株。结论:该方法可作为β-内酰胺类抗生素的常规微生物限度检查方法。 Objective: To establish a limit test method of β-lactam antibiotics. Methods: Antibacterial activity of β-lactam antibiotics was removed by membrane filtration and adding β-lactamase into the medium. Results: The method meets the basic requirements of the 2005 edition of Chinese Pharmacopoeia validation test. Bacillus subtilis was the most sensitive to β-lactam antibiotics among the 5 strains tested, which could be used as the quality control strain of microbial limit of β-lactam antibiotics. Conclusion: This method can be used as a routine microbial limit of β-lactam antibiotics.