Rapid and real-time detection of Porcine Sapelovirus by reverse transcription loop-mediated isothermal amplification assay

被引：7

作者：

Wang, Chunyan ^{[1
]}

Yu, Dayi ^{[2
]}

Cui, Li ^{[1
]}

Hua, Xiuguo ^{[1
]}

Yuan, Congli ^{[1
]}

Sun, Huan ^{[1
]}

Liu, Yuxiao ^{[1
]}

机构：

[1] Shanghai Jiao Tong Univ, Sch Agr & Biol, Shanghai Key Lab Vet Biotechnol, Shanghai 200030, Peoples R China

[2] Anim Dis Control Ctr Minhang Dist, Shanghai, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2014年 / 203卷

关键词：

Porcine Sapelovirus; RT-LAMP; Detection; Picornaviridae; ENTEROVIRUSES; VIRUS;

D O I：

10.1016/j.jviromet.2014.03.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The present study describes the development and validation of a one-step, single-tube, and real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting Porcine Sapelovirus. RT-LAMP characterized by one strand displacement reaction with the specific stem-loop structure and Bst DNA polymerase could be finished in 60 mm under isothermal condition at 63 C. RT-LAMP assay showed higher sensitivity with 10(1) copies/mu L than RT-PCR for the detection of Sapelovirus. The specificity of RT-LAMP assay was validated by the absence of any cross-reaction with other closely related virus in Picornaviridae group and other common virus causing porcine diarrhea. 7 positive Sapelovirus infection out of 63 fecal samples were identified using RT-LAMP, while 5 positive samples were determined by a conventional RT-PCR. A cost-effective method for Saplovirus detection with high sensitivity and specificity was developed and evaluated. (C) 2014 Elsevier B.V. All rights reserved.

引用

页码：5 / 8

页数：4

共 17 条

[1] Complete Genome Sequence of a Novel Porcine Sapelovirus Strain YC2011 Isolated from Piglets with Diarrhea [J].

Chen, Junwei ;

Chen, Feng ;

Zhou, Qingfeng ;

Li, Wei ;

Song, Yanhua ;

Pan, Yongfei ;

Zhang, Xiangbin ;

Xue, Chunyi ;

Bi, Yingzuo ;

Cao, Yongchang .

JOURNAL OF VIROLOGY, 2012, 86 (19) :10898-10898

[2] Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus [J].

Dukes, J. P. ;

King, D. P. ;

Alexandersen, S. .

ARCHIVES OF VIROLOGY, 2006, 151 (06) :1093-1106

[3]

Fauquet C.M., 2005, Virus taxonomy: VIIIth report of the international committee on taxonomy of viruses

[4]

GREIG AS, 1961, CAN J COMP MED VET S, V25, P142

[5]

Jauka T.I., 2010, GENERATION POLYCLONA

[6] Detection of porcine teschoviruses and enteroviruses by LightCycler real-time PCR [J].

Krumbholz, A ;

Wurm, R ;

Scheck, O ;

Birch-Hirschfeld, E ;

Egerer, R ;

Henke, A ;

Wutzler, P ;

Zell, R .

JOURNAL OF VIROLOGICAL METHODS, 2003, 113 (01) :51-63

[7] Sequencing of porcine enterovirus groups II and III reveals unique features of both virus groups [J].

Krumbholz, A ;

Dauber, M ;

Henke, A ;

Birch-Hirschfeld, E ;

Knowles, NJ ;

Stelzner, A ;

Zell, R .

JOURNAL OF VIROLOGY, 2002, 76 (11) :5813-5821

[8] Isolation and characterization of the first Chinese porcine sapelovirus strain [J].

Lan, Daoliang ;

Ji, Wenhui ;

Yang, Shixing ;

Cui, Li ;

Yang, Zhibiao ;

Yuan, Congli ;

Hua, Xiuguo .

ARCHIVES OF VIROLOGY, 2011, 156 (09) :1567-1574

[9] Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hepatitis E Virus [J].

Lan, Xi ;

Yang, Bin ;

Li, Bao Yu ;

Yin, Xiang Ping ;

Li, Xue Rui ;

Liu, Ji Xing .

JOURNAL OF CLINICAL MICROBIOLOGY, 2009, 47 (07) :2304-2306

[10] Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation [J].

Mori, Y ;

Nagamine, K ;

Tomita, N ;

Notomi, T .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2001, 289 (01) :150-154

← 1 2 →