Detection of swine transmissible gastroenteritis coronavirus using loop-mediated isothermal amplification

被引：20

作者：

Chen, Qin ^{[1
]}

Li, Jian ^{[2
]}

Fang, Xue-En ^{[1
]}

Xiong, Wei ^{[2
]}

机构：

[1] Shanghai Univ, Sch Life Sci, Shanghai 200444, Peoples R China

[2] Shanghai Entry Exit Inspect & Quarantine Bur, Shanghai 200135, Peoples R China

来源：

VIROLOGY JOURNAL | 2010年 / 7卷

关键词：

RAPID DETECTION; SYNDROME VIRUS; DISEASE VIRUS; ANTIBODIES; INFECTION; ELISA; DNA;

D O I：

10.1186/1743-422X-7-206

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A conserved nucleic acid fragment of the nucleocapsid gene of Swine Transmissible Gastroenteritis Coronavirus (TGEV) was chosen as the target, six special primers were designed successfully. Loop-mediated isothermal amplification (LAMP) was developed to detect the TGEV by incubation at 60 degrees C for 1 h and the product specificity was confirmed by HphI digestion. Standard curves with high accuracy for TGEV quantization was constructed by adding 1 x SYBR greenI in the LAMP reaction. The assay established in this study was found to detect only the TGEV and no cross-reaction with other viruses, demonstrating its high specificity. By using serial sample dilutions as templates, the detection limit of LAMP was about 10 pg RNA, 10 times more sensitive than that of PCR and could be comparable to the nest-PCR.

引用

页数：5

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