猪鼻支原体LAMP快速诊断检测方法的建立
发布时间：2019-02-21 17:34
【摘要】：采用环介导等温扩增(LAMP)技术,以猪鼻支原体的P37基因序列为靶基因,设计1组特异性引物,对反应体系的引物浓度、MgSO_4、dNTPs、反应温度和时间进行优化,并进行了特异性和灵敏度试验,旨在建立猪鼻支原体(Mhr)的简便、快速、准确的分子检测方法。同时应用所建立的方法对临床样品进行检测,结果显示,最佳反应体系为MgSO_46 mmol/L,dNTPs 1.2 mmol/L,内引物(FIP/BIP)1.6 μmol/L,外引物(F3/B3)0.2 μmol/L,Bst DNA聚合酶8 U,60℃扩增60 min。灵敏度是常规PCR的100倍并且与其他病原菌无任何交叉反应。对23份呼吸道疾病临床样品进行检测的结果显示,普通PCR和LAMP的检出率分别为23.4%和79.6%。表明建立的LAMP检测方法可以用于Mhr的常规检疫。
[Abstract]:A group of specific primers were designed using the sequence of P37 gene of Mycoplasma suis as target gene. The concentration of primers, the temperature and time of MgSO_4,dNTPs, reaction were optimized. The specificity and sensitivity tests were carried out to establish a simple, rapid and accurate molecular detection method for Mycoplasma suis (Mhr). The results showed that the best reaction system was MgSO_46 mmol/L,dNTPs 1.2 mmol/L, internal primer (FIP/BIP) 1.6 渭 mol/L, external primer (F3/B3) 0.2 渭 mol/L,. Amplification of Bst DNA Polymerase 8U at 60 鈩，
本文编号：2427726
[Abstract]:A group of specific primers were designed using the sequence of P37 gene of Mycoplasma suis as target gene. The concentration of primers, the temperature and time of MgSO_4,dNTPs, reaction were optimized. The specificity and sensitivity tests were carried out to establish a simple, rapid and accurate molecular detection method for Mycoplasma suis (Mhr). The results showed that the best reaction system was MgSO_46 mmol/L,dNTPs 1.2 mmol/L, internal primer (FIP/BIP) 1.6 渭 mol/L, external primer (F3/B3) 0.2 渭 mol/L,. Amplification of Bst DNA Polymerase 8U at 60 鈩，
本文编号：2427726
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