16S rDNA序列分析法在注射剂微生物污染鉴定及溯源分析中的应用

Identification and homology analysis of microbial contamination in injection by 16S rDNA sequence analysis

目的对一次注射剂无菌检查阳性结果进行溯源。方法采用16S rDNA序列分析法对污染菌A1和污染调查中采集的16株葡萄球菌进行鉴定和同源性分析,并将分析结果用脉冲场凝胶电泳进行验证。结果A1的鉴定结果为科氏葡萄球菌;科氏葡萄球菌科氏亚种菌株的核糖体16S亚基序列完全相同;结合PFGE分析显示A1来自无菌检查使用的隔离器污染。结论制药企业可以使用16S rDNA序列分析法进行生产和检验中污染菌的鉴定和溯源,但对于与如科氏葡萄球菌科氏亚种一样核糖体16S亚基序列完全相同的菌株,需要结合其他同源性分析方法进行最终的判定。

Objective To trace the source of a positive result from injection sterility test.Methods Positive A1 and 16 Staphylococcus collected in pollution investigation were identified and applied homology analysis by 16S rDNA sequence analysis,and the results were verified by pulsed field gel electrophoresis(PFGE).Result A1 was identified as Staphylococcus cohnii,and the 16S rDNA sequence of Staphylococcus cohnii ssp.Cohnii was exactly identical.PFGE analysis showed that A1 was generated from the isolator system used in sterility testing.Conclusion 16S rDNA sequence analysis can be used to identify and trace the source of microbial contamination.However,for strains with the identical 16S rDNA sequence as Staphylococcus cohnii ssp.Cohnii,it is necessary to apply other homology analysis methods to draw the final conclusion.