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[目的]建立环介质等温扩增技术(LAMP),快速检测恙虫病东方体。[方法]根据staM56KD型特异抗原基因序列,用生物软件设计4条LAMP检测特异性引物,优化反应条件。[结果]当反应体系为25μl,内外引物浓度比1︰4,95℃变性5min,60℃等温扩增1h,80℃2min终止反应,扩增出梯形带,效果最佳。[结论]环介质等温扩增技术能够快速检测恙虫病东方体,检出率与PCR法相近。
[Objective] To establish isothermal amplification of loop medium (LAMP) for the rapid detection of Orientia tsutsugamushi. [Method] According to the sequence of staM56KD specific antigen, 4 LAMP specific primers were designed by biological software to optimize the reaction conditions. [Result] When the reaction system was 25μl, the internal and external primer concentration was 1: 4, the denaturation at 95 ℃ for 5min, the isothermal amplification at 60 ℃ for 1h, the reaction was stopped at 80 ℃ for 2min, and the trapezoidal band was amplified. [Conclusion] The isothermal amplification of the circular medium can detect Orientia tsutsugamushi fast, the detection rate is close to that of PCR method.