一种用于检测恶性疟原虫的双环引物环介导等温扩增方法  

Establishment of Two Pairs of Loop Primers Involved LAMP Method to Detect Plasmodium falciparum

作　　者：吕沁风[1] 姜侃 杨永耀[1] 何蕾[1] 罗鹏[1] 吴忠华[1] 

机构地区：[1]浙江国际旅行卫生保健中心,浙江杭州310003 [2]浙江省质量监督检测研究院,浙江杭州310003

出　　处：《生物技术通讯》2015年第5期712-715,共4页Letters in Biotechnology

基　　金：国家质检总局科技计划(2013IK245);质检公益性行业科研专项(201010043)

摘　　要：目的:设计一种快速环介导等温扩增(LAMP)检测方法,用于恶性疟原虫的检测。方法:根据恶性疟原虫18S r RNA基因设计一组引物,包括外引物、内引物、环引物及第二对环引物,用于恶性疟原虫的LAMP检测。结果:双环引物LAMP检测法灵敏度可达10拷贝DNA模板/μL,可特异地检测出恶性疟原虫DNA,3次重复试验Ct值的CV小于5%,比普通加环引物LAMP检测法时间缩短10 min左右,提高了反应的检测效率。结论:建立的双环引物LAMP方法是一种高效、快速的检测方法。Objective： To establish a rapid loop-mediated isothermal amplification（LAMP） detection method of Plasmodium falciparum. Methods： A set of primers including the outer primers, internal primers, loop primers and the second loop primers were designed according to gene of 18S rRNA of P.falciparum. Results： The sensitivity of this LAMP detection method reached 1×10 copies of template/μL, and the specificity for P.falciparum was veri- fied. The CV of Ct value of three repeated experiments was lower than 5%. More importantly, this two pairs of loop primers involved LAMP method saved 10 minutes compared with conventional LAMP method. Conclusion： The second pair of loop primer is proved to increase the isothermal amplification loop rate, and this established LAMP method is an efficient and reliable detection method of P.falciparum.

关 键 词：环介导等温扩增 双环引物 恶性疟原虫 检测 

分 类 号：Q78[生物学—分子生物学] R531.3[医药卫生—内科学]