NaHCO_3缓冲体系的动态变化对培养法支原体检查的干扰作用  

Dynamic change of sodium bicarbonate buffer system interferences in mycoplasma examination by culture

作　　者：刘辉[1] 李婷婷[1] 邱璐[1] 张波[1] 崔越[1] 戴长河 林洪娟[1] 夏青娟[1] 惠琦[1] 杨红育[1] 

机构地区：[1]长春生物制品研究所有限责任公司,吉林长春130062 [2]长春祈健生物制品有限公司,吉林长春130012

出　　处：《中国生物制品学杂志》2014年第12期1543-1546,共4页Chinese Journal of Biologicals

摘　　要：目的探讨NaHCO3缓冲体系的动态变化对培养法支原体检查的干扰作用,确保生产过程中支原体污染检查的准确性。方法用NaHCO3缓冲液和NaOH分别调整狂犬病病毒收获液及不含病毒样本的病毒稀释液p H至6.92、7.51、7.65、7.85、8.04和8.48,分别接种于支原体半流体培养基和精氨酸支原体肉汤培养基,进行培养法支原体检查,检测结果为阳性的样品分别用指示细胞法(DNA染色法)和实时荧光PCR法进行复检。结果用NaHCO3缓冲液调整狂犬病病毒收获液p H值至7.85以上时,培养法支原体检查结果为阳性,但指示细胞法(DNA染色法)和实时荧光PCR法复检结果均为阴性,排除样品污染支原体的可能性。结论用NaHCO3调整病毒培养物等生物样品p H值达7.85以上时,对培养法支原体检测结果产生干扰作用。Objective To investigate the interference of dynamic change of sodium bicarbonate buffer system to mycoplasma examination by culture so as to ensure the accuracy of inspection on mycoplasma contamination. Methods The p H values of rabies virus harvest were adjusted with sodium bicarbonate buffer to 6. 92,7. 51,7. 65,7. 85,8. 04 and 8. 48 respectively,using the diluent containing no virus as control,then inoculated to semi-fluid and arginine broth media respectively for examination of mycoplasma. The samples with positive results were checked by DNA staining and real-time fluorescent PCR. Results When the p H value of rabies virus harvest was adjusted with sodium bicarbonate buffer to more than 7. 85,the examination result of mycoplasma by culture was positive,while those by DNA staining and real-time fluorescent PCR were negative,which excluded the possibility of mycoplasma contamination in the samples.Conclusion The adjusting of p H value of samples with sodium bicarbonate buffer to more than 7. 85 might interfere in the examination result of mycoplasma by culture.

关 键 词：支原体 NAHCO3 培养法 

分 类 号：R375[医药卫生—病原生物学] R392-33[医药卫生—基础医学]