[1]张玉霞,杨金兴,孟 凯,等.鹅星状病毒LAMP快速检测方法的建立与应用[J].中国预防兽医学报,2019,(11):1137-1140.[doi:0.3969/j.issn.1008-0589.201812039]
 ZHANG Yu-xia,YANG Jin-xing,MENG Kai,et al. Establishment and application of LAMP method for rapid detection of goose astrovirus[J].Chinese journal of preventive veterinary medicine,2019,(11):1137-1140.[doi:0.3969/j.issn.1008-0589.201812039]
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鹅星状病毒LAMP快速检测方法的建立与应用()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年11
页码:
1137-1140
栏目:
诊断和检测技术
出版日期:
2019-12-25

文章信息/Info

Title:
 

Establishment and application of LAMP method for rapid detection of goose astrovirus

文章编号:
1008-0589(2019)11-1137-04
作者:
 

张玉霞1杨金兴1孟 凯1蔡李萌2袁小远1*王友令1*

 (1. 山东省农业科学院 家禽研究所,山东 济南 250100;2. 青岛农业大学 动物医学院,山东 青岛 266109)
Author(s):
 

ZHANG Yu-xia1 YANG Jin-xing1 MENG Kai1 CAI Li-meng2 YUAN Xiao-yuan1* WANG You-ling1*

 (1. Institute of Poultry Science, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
2. College of veterinary medicine, Qingdao Agricultural University, Qingdao 266109, China)
关键词:
鹅星状病毒LAMP快速检测RT-PCR
Keywords:
goose Astrovirus  LAMP  rapid detection  RT-PCR
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201812039
文献标志码:
A
摘要:
鹅星状病毒是一种新发的可导致雏鹅以痛风为主要特征的传染性病原。为建立鹅星状病毒(AstV)的快速检测方法,本研究根据鹅AstV ORF1b基因的保守序列设计了一组特异性的LAMP引物,建立鹅AstV的LAMP快速检测方法,并检测其特异性和灵敏度。结果显示:建立的LAMP方法的最佳反应温度为64 ℃;仅能扩增鹅AstV,而对鹅细小病毒、鹅副黏病毒、鸭坦布苏病毒及鹅源H9N2亚型禽流感病毒均不能扩增;能够检测到的最低模板(cDNA)浓度为1 ng/μL,因此该方法具有较好的特异性和敏感性。临床样品检测结果显示:建立的LAMP方法与RT-PCR方法阳性符合率为95.65 %。综上,本研究建立的LAMP方法能够特异、快速地检测新发鹅AstV,为养殖基地、科研和检验检疫部门提供了一种快速检测该病原的技术手段。
Abstract:
Goose Astrovirus is a newly emerging infectious agent that mainly causes gout in goslings. In order to establish a rapid assay for the detection of goose Astrovirus, we designed a set of LAMP primers specificlly targeting the conserved regions of ORF1b gene and established a loop-mediated isothermal amplification (LAMP) method. The specificity and sensitivity of this LAMP assay for the detection of Astrovirus were evaluated. The results showed that the established LAMP assay was sensitive and specific at 64℃, since goose Astrovirus could be amplified by this method with a detection limit of 1 ng/μL of cDNA, while neither goose parvovirus, goose paramyxovirus, duck Tembusu virus, nor goose H9N2 subtype influenza virus was amplified by this method. With the use of clinical sample positive for goose Astrovirus, this LAMP method yield an agreement of 95.65% with RT-PCR test. In conclusion, the present study estabulished a LAMP assay for specific and rapid detection of goose Astrovirus, which provides a quick diagnostic method of Astrovirus infection for breeding bases, scientific research institutions, and departments of inspection and quarantine.

参考文献/References:

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(本文编辑:李 爽;英文编辑:李 海)

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更新日期/Last Update: 2019-12-09