| 寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用 |
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| 引用本文： | 于宁,刘宇梦,李成辉,李卓昕,汪伟,孙文超,鲁会军,金宁一.寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用[J].中国动物传染病学报,2021(1). |
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| 作者姓名： | 于宁 刘宇梦 李成辉 李卓昕 汪伟 孙文超 鲁会军 金宁一 |
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| 作者单位： | 军事科学院军事兽医研究所;延边大学农学院;广西大学动物科学技术学院;温州大学病毒学研究所 |
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| 基金项目： | 国家科技重大专项(2018ZX10101003,2018ZX10102001);浙江省青年基金项目(LQ19C180001);温州市基础性科研项目(N20190005,N20180010)。 |
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| 摘 要： | 为建立一种针对寨卡病毒的快速诊断方法,本研究根据寨卡病毒的3’端保守基因序列,设计合成1对引物,建立了检测寨卡病毒的荧光定量PCR方法。结果显示:所建立的检测方法的Ct值与标准品在1.41×10^1~1.41×10^10^ copies/μL具有良好的线性关系,相关性为1,斜率为-3.502;灵敏性结果显示,该方法的检测限度为1.41×10^1 copies/μL,是普通PCR的10000倍;特异性结果显示,对CHIKV、DENV和JEV无特异性扩增,特异性强;重复性试验结果显示,组内和组间变异系数均小于1%,重复性好。本研究建立的SYBR Green I real-time PCR检测方法,可用于寨卡病毒感染的快速诊断。
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| 关 键 词： | 寨卡病毒 SYBR GreenⅠreal-time PCR 应用方法 |
Establishment and Application of SYBR GreenⅠReal-Time PCR for the Detection of Zika Virus |
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| YU Ning,LIU Yumeng,LI Chenghui,LI Zhuoxin,WANG Wei,SUN Wenchao,LU Huijun,JIN Ningyi.Establishment and Application of SYBR GreenⅠReal-Time PCR for the Detection of Zika Virus[J].Chinese Journal of Animal Infectious Diseases,2021(1). |
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| Authors: | YU Ning LIU Yumeng LI Chenghui LI Zhuoxin WANG Wei SUN Wenchao LU Huijun JIN Ningyi |
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| Institution: | (l.Institute of Military Veterinary,The Academy of Military Medical Sciences,Changchun 130122,China;Agricultural college,Yanbian University,Yanji 133000,China;Animal Science and Technology,Guangxi University College,Nanning 530001,China;Institute of Virology,Wenzhou University,Wenzhou 325035,China) |
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| Abstract: | To develop a rapid diagnostic method for Zika virus,a pair of specific primers was designed and synthesized based on the 3’-end conserved sequence of Zika virs for the development of a fluorescent quantitative PCR method.The results showed this PCR method had a good linear relationship between the Ct values with the standard template in the range from 1.41×10^1 to 1.41×10^10copies/μL,and the correlation was 1.000 with a slope of-3.502.In addition,the sensitivity testing found the detection limit was 1.41×10^1 copies/μL,which was 10,000 times higher than that of the ordinary PCR.The specificity results revealed that this method had no specific amplification for CHIKV,DENV and JEV.The repeatability experiment showed that the coefficient of variation between and within groups was less than 1%.In conclusion,the SYBR Green I real-time PCR method developed here might be used for rapid diagnosis of Zika virus infection. |
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| Keywords: | Zika virus SYBR GreenⅠreal-time PCR Application |
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