| RT-PCR方法检测单核细胞增生李斯特活菌研究 |
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| 引用本文： | 李善志,吴清平,张菊梅,郭伟鹏. RT-PCR方法检测单核细胞增生李斯特活菌研究[J]. 中国卫生检验杂志, 2006, 16(6): 641-643 |
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| 作者姓名： | 李善志 吴清平 张菊梅 郭伟鹏 |
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| 作者单位： | 1. 广东省微生物研究所,广东省菌种保藏与应用重点实验室,广州,510070;中国科学院研究生院,北京,100049
2. 广东省微生物研究所,广东省菌种保藏与应用重点实验室,广州,510070 |
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| 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1×104cfu/m l,人工污染样品猪肉、虾肉和牛奶等培养12~16 h,可达4.5 cfu/m l(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。
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| 关 键 词： | 单核细胞增生李斯特菌 活菌检测 hlyA基因 |
| 文章编号： | 1004-8685（2006）06-0641-03 |
| 收稿时间： | 2006-01-10 |
| 修稿时间： | 2006-01-10 |
Detection of viable Listeria monocytogenes with RT-PCR assay |
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| Li Shan-zhi,Wu Qing-ping,Zhang Ju-mei,Guo Wei-peng. Detection of viable Listeria monocytogenes with RT-PCR assay[J]. Chinses Journal of Health Laboratory Technology, 2006, 16(6): 641-643 |
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| Authors: | Li Shan-zhi Wu Qing-ping Zhang Ju-mei Guo Wei-peng |
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| Abstract: | Objective: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 palse positive problems. |
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| Keywords: | mRNA RT-PCR |
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