LAMP结合基因芯片技术在快速检测嗜肺军团菌中的应用

A rapid detection of Legionella Pneumophila by LAMP in combination with gene chip

目的:开发一种可以快速、准确检测嗜肺军团菌的检测方法。方法:采用环介导等温核酸扩增技术(loop-mediated isothermal amplification,LAMP)对嗜肺军团菌的特异性DNA保守片段和其他对照菌种DNA进行扩增,将扩增的DNA与基因芯片进行杂交,观察是否可在芯片相应的区域出现杂交信号。结果:只有嗜肺军团菌DNA经LAMP反应后出现特异性的扩增条带,与PCR扩增基因的最低稀释倍数105相比,LAMP最低稀释倍数可达107,反应的灵敏度比PCR高100倍左右,同时将经LAMP扩增后的基因与基因芯片杂交,相应的嗜肺军团菌区域可出现杂交信号。结论:LAMP技术结合基因芯片杂交技术具有灵敏度高,特异性好,简便快速等优势,可提高检验鉴定结果的准确性。

Objective To establish a rapid and accurate method to detect the Legionella Pneumophila（LP）.Methods The loop-mediated isothermal amplification（LAMP） to amplify the specific DNA fragments of LP and the gene chip technology was further used to identify the LP sequence.Results The DNA bands corresponding to LP DNA could be specifically amplified by the LAMP reaction.For normal PCR,the minimum dilution of genomic DNA was 10-5,however,for the LAMP,the minimum dilution of genomic DNA was up to 10-7,so the sensitivity of the LAMP reaction is about 100 times higher than of normal PCR.The combination of the LAMP and gene chip contributed to demonstrating the specific LP sequence signal.Conclusion Combination of LAMP and gene chip is a simple,accutate and rapid technology to detect LP with,high sensitivity and specificity.