摘要
目的:建立单核细胞增生李斯特活菌快速检测技术,解决PCR带来的假阳性问题。方法:采用以mRNA为基础的RT—PCR方法对单核细胞增生李斯特氏菌的hlyA基因进行检测,设计引物,扩增,并作了特异性和灵敏度比较。结果:该引物能较好地扩增单核细胞增生李斯特菌的特异性片断,而对其它的李斯特菌以及常见食源性致病菌无特异性扩增条带;检测的灵敏度纯菌可达到1×10^4cfu/ml,人工污染样品猪肉、虾肉和牛奶等培养12—16h,可达4.5cfu/ml(g)。结论:可以应用该方法对样品中单核细胞增生李斯特活菌进行检测,能较好地解决PCR检测所带来的假阳性问题。
Objectlve:To develop a method for rapid detection of viable Listeria monocytogenes. Methods:The hlyA gene of Listeria monocytogenes was detected by mRNA - based RT - PCR. The sensitivity and specificity of the RT - PCR method were compared to conventional methods. Results: Only special fragment of Listeria monocytogenes could be extended by the primers. There was no crossreaction with Listeria and other food - borne pathogenes. The sensitivity of detection was 4. 5 cfu/ml (g). Conclusion:This method can be used to detect viable Listeria monocytogenes in the samples to solve paise positive problems.
出处
《中国卫生检验杂志》
CAS
2006年第6期641-643,共3页
Chinese Journal of Health Laboratory Technology
基金
广东省科技计划资助项目(2005B10401023)