| 鹅源星状病毒TaqMan荧光定量PCR检测方法的建立及初步应用 |
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| 引用本文： | 苏世博,蒲路莎,陈肖韩,赵丽丽,陈洪岩. 鹅源星状病毒TaqMan荧光定量PCR检测方法的建立及初步应用[J]. 中国比较医学杂志, 2021, 0(3): 43-48 |
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| 作者姓名： | 苏世博 蒲路莎 陈肖韩 赵丽丽 陈洪岩 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所/兽医生物技术国家重点实验室/黑龙江省实验动物与比较医学重点实验室/国家禽类实验动物资源库 |
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| 基金项目： | 国家十三五重点研发计划子课题(2016YFD0500800);黑龙江省青年科学基金项目(QC2018033);黑龙江省自然科学基金重点项目(ZD2016006);中央级公益性科研院所基本科研业务费专项(1610302020008,1610302017013)。 |
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| 摘 要： | 目的 建立一种可快速诊断鹅源星状病毒的荧光定量PCR方法 .方法 分析比对NCBI下载的鹅源星状病毒全基因组序列,在其ORF1a保守区设计一对特异性引物及TaqMan探针,优化反应条件,建立标准曲线,并验证其特异性,敏感性和稳定性.结果 该方法 最低检测线限为1.50×102copies/μL,批内和批间检测变异系数分...
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| 关 键 词： | 鹅源星状病毒 荧光定量PCR 雏鹅痛风 检测方法 |
Establishment and application of TaqMan reverse transcriptionquantitative PCR for newly emerging goose-origin astrovirus in China |
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| SU Shibo,PU Lusha,CHEN Xiaohan,ZHAO Lili,CHEN Hongyan. Establishment and application of TaqMan reverse transcriptionquantitative PCR for newly emerging goose-origin astrovirus in China[J]. Chinese Journal of Comparative Medicine, 2021, 0(3): 43-48 |
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| Authors: | SU Shibo PU Lusha CHEN Xiaohan ZHAO Lili CHEN Hongyan |
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| Affiliation: | (Harbin Veterinary Research Institute,Chinese Academy of Agriculture Sciences,State Key Laboratory of Veterinary Biotechnology,Heilongjiang Provincial Key Laboratory of Laboratory Animal and Comparative Medicine,National Poultry Laboratory Animal Resource Center,Harbin 150069,China) |
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| Abstract: | Objective In this study,we established a fluorescence quantitative PCR method for rapidly diagnosing a new type of goose-origin astrovirus(Go Ast V).Methods After analyzing and comparing the whole-genome sequences of Go Ast V downloaded from NCBI,a pair of specific primers and Taq Man probes were designed in its open reading frame-1 a conservative region.By optimizing the reaction conditions,a standard curve was established,and the specificity,sensitivity and stability were verified.Results The minimum sample size was 1.5×102 copies/μL.The coefficients of variation for the intra-assay and inter-assay detection were<0.5%and 4%,respectively,with no specific amplification of other common waterfowl viruses.The clinical application result showed that the detection rate was significantly higher than that of conventional real-time PCR.Conclusions This fluorescence quantitative PCR method exhibited stronger specificity,higher sensitivity and better reproducibility for early detection and quantitative analysis of Go Ast V. |
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| Keywords: | goose-origin astrovirus RT-qPCR gosling gout detection method |
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