| 检测猪粪便中基因4型戊型肝炎病毒的实时荧光定量RT-PCR方法的建立 |
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| 引用本文： | 贾鹏,金宁一,李霄,朱光泽,刘妍,高鹏,徐晓红,杨恩成,孟日增,阚式绂. 检测猪粪便中基因4型戊型肝炎病毒的实时荧光定量RT-PCR方法的建立[J]. 病毒学报, 2010, 26(1): 33-39 |
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| 作者姓名： | 贾鹏 金宁一 李霄 朱光泽 刘妍 高鹏 徐晓红 杨恩成 孟日增 阚式绂 |
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| 作者单位： | 军事医学科学院,全军基因工程重点实验室,长春,130062;吉林大学,长春,130062;军事医学科学院,全军基因工程重点实验室,长春,130062;长春中医药大学,长春,130021;吉林省进出口检验检疫局,长春,130062 |
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| 基金项目： | 国家自然科学基金(30771609) |
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| 摘 要： | 针对基因4型戊型肝炎病毒(Hepatitis E virus,HEV)基因组ORF3设计特异性引物和探针,建立了一套TaqMan实时荧光定量RT-PCR方法。评价了该方法的准确性和灵敏性,同时与常规RT-PCR和巢式RT-PCR方法进行了对比分析。结果表明,生成的标准曲线显示各浓度范围内具有良好的线性关系,相关系数(r2)为0.994,斜率为-3.312,PCR效率为100%。组内和组间重复性试验相同稀释度标准质粒Ct值之间差异均不显著,并且能够准确的从HEV阳性猪粪便样品中检测到基因4型HEV RNA,具有较好准确性。可检测到戊型肝炎病毒数量为(1.7×101)copies,比普通RT-PCR的灵敏度高100倍,比Nested RT-PCR高10倍。
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| 关 键 词： | 戊型肝炎病毒 RT-PCR 基因4型 |
Establishment of a Fluorescent Real-time Quantitative RT-PCR Assay for Detection of Genotype 4 Hepatitis E Virus in Swine Stools |
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| JIA Peng,,JIN Ning-yi,LI Xiao,ZHU Guang-ze,,LIU Yan,GAO Peng,XU Xiao-hong,YANG En-cheng,MENG Ri-zeng,KAN Shi-fu. Establishment of a Fluorescent Real-time Quantitative RT-PCR Assay for Detection of Genotype 4 Hepatitis E Virus in Swine Stools[J]. Chinese journal of virology, 2010, 26(1): 33-39 |
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| Authors: | JIA Peng JIN Ning-yi LI Xiao ZHU Guang-ze LIU Yan GAO Peng XU Xiao-hong YANG En-cheng MENG Ri-zeng KAN Shi-fu |
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| Affiliation: | 1.Genetic Engineering Laboratory of PLA;Academy of Military Medical Sciences of PLA;Changchun 130062;China;2.Faculty of Agriculture Sciences of Jilin University;3.The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine;Changchun 130021;4.JiLin Entry-Exit Inspection and Quarantine Bureau;China |
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| Abstract: | The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence(swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV.A rapid,sensitive and stable TaqMan Real-time RT-PCR assay was established,and its specificity and sensitivity were assessed,and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed.The results found that the crossing points showed linearly proportional to the logarithm of the input copy number.The correlati... |
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| Keywords: | HEV Real-time RT-PCR genotype 4 |
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