基于微流控芯片技术的五种啮齿类动物疫病检测方法的建立

Development of a Detection Method for Five Rodent Diseases Based on Microfluidic Chip Technology

为提升啮齿类实验动物的检疫效率,本研究拟建立鼠痘病毒、流行性出血热病毒、淋巴细胞性脉络丛脑膜炎病毒、仙台病毒和鼠肝炎病毒联检的微流控芯片方法并进行评价。针对5种病毒基因保守序列设计特异性LAMP引物,将引物包埋于芯片反应槽内,通过检测5种啮齿类动物病毒和另外3种病毒的样本来分析本方法的特异性,通过检测不同浓度的模板来评价本方法的灵敏度和重复性,通过检测35份小鼠的临床阳性样本和30份健康小鼠的肝组织样本来评价本方法的临床应用效果。结果表明,本研究建立的微流控芯片方法特异性好、灵敏度高、重复性佳,各指标间无相互干扰。本方法可提高啮齿类实验动物的检疫效率。

In order to improve the quarantine efficiency of rodent laboratory animals, the present study aimed to develop a microfluidic chip method for detection of Ectromelia virus, Epidemic hemorrhagic fever virus, Lymphocytic choriomeningitis virus, Sendai virus and murine hepatitis virus based on loop-mediate isothermal amplification. Specific primers were designed for the conserved sequences of these viruses, respectively. The primers were embedded in the chip reaction holes. The specificity of this method was analyzed by detecting these 5 rodent viruses and 3 other viruses. The sensitivity and repeatability of the method were evaluated by detecting the templates with different concentrations. 35 clinical positive mouse samples and 30 liver tissue samples of healthy mice were detected to evaluate its application. The results showed that the microfluidic chip method developed in this study had high specificity, sensitivity and repeatability, and no mutual interference among these 5 infectious diseases, indicating that this method could improve the quarantine efficiency of rodent laboratory animals.