Development of a minor groove binder assay for real-time PCR detection of porcine Sapelovirus

被引：11

作者：

Chen, Junwei ^{[1
]}

Chen, Feng ^{[2
]}

Zhou, Qingfeng ^{[3
]}

Li, Wei ^{[3
]}

Chen, Yanshan ^{[3
]}

Song, Yanhua ^{[3
]}

Zhang, Xiangbin ^{[2
]}

Xue, Chunyi ^{[1
]}

Bi, Yingzuo ^{[2
]}

Cao, Yongchang ^{[1
]}

机构：

[1] Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510006, Guangdong, Peoples R China

[2] South China Agr Univ, Coll Anim Sci, Guangzhou 510642, Guangdong, Peoples R China

[3] Guangdong Wens Grp Acad, Guangdong Wens Foodstuffs Grp Co Ltd, Xinxing, Guangdong, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2014年 / 198卷

关键词：

Porcine Sapelovirus; Real-time PCR; Minor groove binder probe; Detection; COMPLETE GENOME SEQUENCE; FECAL SAMPLES; TESCHOVIRUSES; PIGLETS;

D O I：

10.1016/j.jviromet.2013.12.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A 5' conjugated minor groove binder (MGB) probe real-time PCR assay was developed in this study for porcine sapelovirus (PSV) detection and quantitation. Two primers and a MGB probe for the 5' untranslated region (UTR) gene were designed. The assay was capable of detecting about 103 copies/mu l of standard template per reaction. Moreover, it does not detect any of the other RNA viruses that cause diarrhea disease in pigs. The coefficients of variation of intra- and inter-assay reproducibility were both lower than 2%. In 73 field fecal samples, PSV was detected in 46 samples using real-time PCR assay and only 32 samples with a conventional PCR assay. Therefore, the availability of this assay will facilitate further studies on the epidemiology of PSV infection and its role in swine disease. (C) 2013 Elsevier B.V. All rights reserved.

引用

页码：69 / 74

页数：6

共 22 条

[1] A novel sapelovirus-like virus isolation from wild boar [J].

Abe, Masako ;

Ito, Naoto ;

Sakai, Kouji ;

Kaku, Yoshihiro ;

Oba, Mami ;

Nishimura, Miho ;

Kurane, Ichiro ;

Saijo, Masayuki ;

Morikawa, Shigeru ;

Sugiyama, Makoto ;

Mizutani, Tetsuya .

VIRUS GENES, 2011, 43 (02) :243-248

[2] Minor groove binder-conjugated DNA probes for quantitative DNA detection by hybridization-triggered fluorescence [J].

Afonina, IA ;

Reed, MW ;

Lusby, E ;

Shishkina, IG ;

Belousov, YS .

BIOTECHNIQUES, 2002, 32 (04) :940-+

[3] A survey of porcine picornaviruses and adenoviruses in fecal samples in Spain [J].

Buitrago, Dolores ;

Cano-Gomez, Cristina ;

Agueero, Montserrat ;

Fernandez-Pacheco, Paloma ;

Gomez-Tejedor, Concepcion ;

Angel Jimenez-Clavero, Miguel .

JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION, 2010, 22 (05) :763-766

[4] Teschoviruses and sapeloviruses in faecal samples from wild boar in Spain [J].

Cano-Gomez, Cristina ;

Ana Garcia-Casado, Maria ;

Soriguer, Ramon ;

Palero, Ferran ;

Angel Jimenez-Clavero, Miguel .

VETERINARY MICROBIOLOGY, 2013, 165 (1-2) :115-122

[5] Evaluation of a fluorogenic real-time reverse transcription-polymerase chain reaction method for the specific detection of all known serotypes of porcine teschoviruses [J].

Cano-Gomez, Cristina ;

Buitrago, Dolores ;

Fernandez-Pinero, Jovita ;

Fernandez-Pacheco, Paloma ;

Mansilla, Carmen ;

Agueero, Montserrat ;

Angel Jimenez-Clavero, Miguel .

JOURNAL OF VIROLOGICAL METHODS, 2011, 176 (1-2) :131-134

[6] 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

[7] Recombination patterns in aphthoviruses mirror those found in other picornaviruses [J].

Heath, Livio ;

van der Walt, Eric ;

Varsani, Arvind ;

Martin, Darren P. .

JOURNAL OF VIROLOGY, 2006, 80 (23) :11827-11832

[8] A review of RT-PCR technologies used in veterinary virology and disease control: Sensitive and specific diagnosis of five livestock diseases notifiable to the World Organisation for Animal Health [J].

Hoffmann, Bernd ;

Beer, Martin ;

Reid, Scott M. ;

Mertens, Peter ;

Oura, Chris A. L. ;

van Rijn, Piet A. ;

Slomka, Marek J. ;

Banks, Jill ;

Brown, Ian H. ;

Alexander, Dennis J. ;

King, Donald P. .

VETERINARY MICROBIOLOGY, 2009, 139 (1-2) :1-23

[9] Genetic reclassification of porcine enteroviruses [J].

Kaku, Y ;

Sarai, A ;

Murakami, Y .

JOURNAL OF GENERAL VIROLOGY, 2001, 82 :417-424

[10] 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

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