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目的建立一种快速、灵敏、高效的人冠状病毒HKU1(HCoV-HKU1)LAMP扩增检测方法。方法应用生物信息学软件设计5套LAMP扩增引物,以合成的HCoV-HKU1质粒为模板,通过实时浊度法进行最佳引物组的筛选,并分析该方法的特异性和灵敏度。结果 HCoV-HKU1 LAMP扩增方法针对其他病原微生物及人类基因组无反应信号。该方法可以检测到1拷贝/μl的HCoV-HKU1质粒。结论 HCoV-HKU1 LAMP扩增方法特异性好、灵敏度高,在临床鉴别诊断和口岸冠状病毒的检测中具有很好的应用前景。
Objective To establish a rapid, sensitive and efficient method for detecting human coronavirus HKU1 (HCoV-HKU1) LAMP amplification. Methods Five sets of LAMP amplification primers were designed by bioinformatics software. The optimal HCoV-HKU1 plasmid was used as a template to screen the best primer set by real-time turbidimetric method. The specificity and sensitivity of the method were analyzed. Results The HCoV-HKU1 LAMP amplification method showed no response to other pathogenic microorganisms and the human genome. This method can detect 1 copy / μl of HCoV-HKU1 plasmid. Conclusion The method of HCoV-HKU1 LAMP amplification has good specificity and high sensitivity, and has good application prospect in the differential diagnosis of clinical and oral coronavirus.