一次性使用防疫用品中五种控制菌多重PCR检测方法的建立  

Establishment of Multiplex PCR Detection Method for Five Control Bacteria in Disposable Epidemic Prevention Supplies

作　　者：刘欢 聂晶 王典 彭彩娟 王烨婷 黄河 LIU Huan;NIE Jing;WANG Dian;PENG Cai-juan;WANG Ye-ting;HUANG He(Hubei Medical Devices Quality Supervision and Test Institute,Hubei Wuhan 430000)

机构地区：[1]湖北省医疗器械质量监督检验研究院,湖北武汉430000

出　　处：《中国医疗器械信息》2025年第1期53-56,共4页China Medical Device Information

摘　　要：目的:建立一次性使用防疫用品中控制菌检测的多重PCR方法。方法:根据金黄色葡萄球菌Nuc、铜绿假单胞菌oprL、大肠埃希菌phoA、溶血性链球菌Mf、白色念珠菌SAP基因的保守序列设计引物,建立多重PCR反应体系,并验证其特异性、敏感性和重复性。结果:多重PCR扩增Nuc、oprL、phoA、Mf以及SAP基因的片段大小依次为486、305、666、808、265bp;检出限依次为2.3×10^(3)CFU/mL、2.0×10^(3)CFU/mL、1.1×10^(3)CFU/mL、7.0×10^(2)CFU/mL、4.1×10^(3)CFU/mL。结论:本研究建立的多重PCR检测方法能同时检测一次性防疫用品中的五种控制菌,方法灵敏、特异且重复性良好,能有效缩短检测周期,提高检验检测效率。Objective:To establish a multiplex PCR method for detection of control bacteria in the disposable epidemic prevention supplies.Methods:According to the conserved sequences of staphylococcus aureus Nuc,pseudomonas aeruginosa oprL,escherichia coli phoA,hemolytic streptococcus Mf and candida albicans SAP genes,the corresponding primers were designed to establish a multiplex PCR reaction system.The specificity,sensitivity and repeatability of the method were tested.Results:The fragment sizes of Nuc,oprL,phoA,Mf and SAP genes were 486 bp,305 bp,666 bp,808 bp and 265 bp,respectively.The corresponding detection limits were 2.3×10^(3)CFU/mL,2.0×10^(3)CFU/mL,1.1×10^(3)CFU/mL,7.0×10^(2)CFU/mL,4.1×10^(3)CFU/mL,respectively.Conclusion:The multiplex PCR detection method established in this study can simultaneously detect the five control bacteria in disposable epidemic prevention supplies.The method is sensitive,specific and reproducible,and can effectively shorten the detection cycle and improve the detection efficiency.

关 键 词：一次性使用防疫用品 微生物限度 控制菌检查 多重PCR 

分 类 号：TH771[机械工程—仪器科学与技术]