A real-time PCR method for detection of porcine circovirus 2

YU Jing; LAO Xiujie; CHEN Yanyong; HE Xiaojiang; DAI Bing; ZHAO Ayong; WANG Xiaodu; SONG Houhui

doi:10.11833/j.issn.2095-0756.2016.02.023

Volume 33 Issue 2

Mar. 2016

Turn off MathJax

Article Contents

Article Navigation > Journal of Zhejiang A&F University > 2016 > 33(2): 357-363

YU Jing, LAO Xiujie, CHEN Yanyong, HE Xiaojiang, DAI Bing, ZHAO Ayong, WANG Xiaodu, SONG Houhui. A real-time PCR method for detection of porcine circovirus 2[J]. Journal of Zhejiang A&F University, 2016, 33(2): 357-363. doi: 10.11833/j.issn.2095-0756.2016.02.023

Citation:

YU Jing, LAO Xiujie, CHEN Yanyong, HE Xiaojiang, DAI Bing, ZHAO Ayong, WANG Xiaodu, SONG Houhui. A real-time PCR method for detection of porcine circovirus 2[J]. Journal of Zhejiang A&F University, 2016, 33(2): 357-363. doi: 10.11833/j.issn.2095-0756.2016.02.023

A real-time PCR method for detection of porcine circovirus 2

doi: 10.11833/j.issn.2095-0756.2016.02.023

1.
College of Animal Science and Technology, Zhejiang A & F University, Lin’an 311300, Zhejiang, China

Funds:

浙江省科技厅公益项目（2014C32061）;浙江农林大学人才启动项目（2011FR025，2013FR077）;浙江省自然科学基金资助项目（LQ14C010007）;浙江农林大学大学生科技创新训练计划项目（201301017）;浙江农林大学面上基金项目（2013FK001）

Received Date: 2015-04-19
Rev Recd Date: 2015-04-19
Publish Date: 2016-04-20

Abstract

To develop an accurate and rapid detection method for disease screening and prevention with porcine circovirus type 2 （PCV2）, a single-strand DNA virus that infects pigs, primers targeted to the ORF2 fragment of the PCV2 conserved region were designed. A real time polymerase chain reaction （PCR） method was developed using SYBR Green as a fluorescent dye and serial dilutions of ORF2 recombinant plasmid to construct a standard curve for an absolute quantification. Results showed that the detection limit was obtained with 101 copy DNA per microliter. This method exhibited 1 000 times higher sensitivity than conventional PCR, only specific identification of PCV2, and better repeatability with the less than 2% variation coefficient of intra- or inter-assay experiments. A total of 34 PCV2 positive clinical samples were confirmed using this real time PCR method, demonstrating 100% agreement in comparison with the conventional PCR having only 50% agreement. This real time PCR method could provide a valuable tool for PCV2 prevention and control.［Ch, 4 fig. 4 tab. 26 ref.］
- zoology,
- porcine,
- porcine circovirus 2,
- real-time PCR

Relative Article

[1]	ZHAO Yu, LIN Lin, WANG Qun, ZHANG Guozhe, WANG Jie, SHANG Linxue, HONG Sidan, MA Qingqing, GU Cuihua. Screening and validation of reference genes in Heimia myrtifolia in different tissues and under drought stress . Journal of Zhejiang A&F University, 2023, 40(3): 665-672. doi: 10.11833/j.issn.2095-0756.20220342
[2]	YE Bihuan, CHEN Youwu, SHU Jinping, ZHANG Wei, ZHANG Yabo, LI Haibo, SONG Qiyan. Screening and application of reference genes for qRT-PCR in bamboo wireworm . Journal of Zhejiang A&F University, 2021, 38(3): 644-651. doi: 10.11833/j.issn.2095-0756.20200492
[3]	SHI Lin, WU Yuan, BA Shaobo, LIU Zhengkui, CHEN Lin, WANG Lei, SHAO Chunyan, SUN Jing, ZHOU Yingshan, WANG Xiaodu, SONG Houhui. Development of Taqman real time fluorescent quantitative PCR for porcine circovirus type 2 . Journal of Zhejiang A&F University, 2018, 35(6): 1133-1138. doi: 10.11833/j.issn.2095-0756.2018.06.018
[4]	WANG Yali, HE Ke, YU Jing, YANG Songbai, ZHAO Ayong. Prediction and validation of miRNA targeting the porcine CLTC gene . Journal of Zhejiang A&F University, 2017, 34(3): 389-394. doi: 10.11833/j.issn.2095-0756.2017.03.002
[5]	LI Yacang, FENG Zhongke, HUANG Jixia, YANG Liu. Habitat suitability evaluation for Amur tigers in northeast China based on GIS and RS . Journal of Zhejiang A&F University, 2016, 33(2): 265-271. doi: 10.11833/j.issn.2095-0756.2016.02.011
[6]	ZHU Zhixiang, SHI Haojie, LEI Feibin, ZHANG Chuanqing. Quantifying Botryosphaeria dothidea infection causing canker disease on Carya cathayensis using real-time PCR . Journal of Zhejiang A&F University, 2016, 33(2): 364-368. doi: 10.11833/j.issn.2095-0756.2016.02.024
[7]	YANG Xiaolan, GONG Hede. Seed predation by rodents in northwest Yunnan Province . Journal of Zhejiang A&F University, 2015, 32(3): 440-445. doi: 10.11833/j.issn.2095-0756.2015.03.017
[8]	GUO Shuai, XU Qiufang, SHEN Zhenming, LI Songhao, QIN Hua, LI Yongchun. Response of soil ammonia-oxidizing organisms on fertilization and mulch in Phyllostachys violascens stands . Journal of Zhejiang A&F University, 2014, 31(3): 343-351. doi: 10.11833/j.issn.2095-0756.2014.03.003
[9]	SHEN Hongxia, HAN Xiujie, ZHAO Fanfan, ZHANG Baoxin, YU Fengyan, WANG Xiaodu. Expression and antibody preparation of nonstructural protein 1 for Japanese encephalitis virus from pigs . Journal of Zhejiang A&F University, 2013, 30(3): 396-400. doi: 10.11833/j.issn.2095-0756.2013.03.015
[10]	YU Yun-wei, YING Ye-qing, REN Li-ping, HU Jia-fu, ZHAO A-yong. Community structure and biodiversity of soil animals in bamboo stands of Lin’an，Zhejiang . Journal of Zhejiang A&F University, 2012, 29(4): 581-587. doi: 10.11833/j.issn.2095-0756.2012.04.015
[11]	WANG Xiao-du, ZHAO A-yong, DENG Xu-fang, MA Zhi-yong. Expression and activity analysis of mouse mdm2 . Journal of Zhejiang A&F University, 2012, 29(2): 155-160. doi: 10.11833/j.issn.2095-0756.2012.02.001
[12]	WANG Rong. Principal component analysis and evaluation of waterfowl habitats along the coastline of Fujian Province . Journal of Zhejiang A&F University, 2011, 28(3): 472-478. doi: 10.11833/j.issn.2095-0756.2011.03.020
[13]	ZHU Xi, WANG Qing-liang, ZHAN Yin-bo, HAN Pu, JIANG Hong-yun, ZHOU Wen, HE Yu-bo. Fauna and distribution of amphibian and reptile on Putuoshan Island，Zhejiang . Journal of Zhejiang A&F University, 2009, 26(5): 708-713.
[14]	ZHU Gui-shou, DING Liang-dong, YU Gen-lian, ZHOU Xiao-li. Wildlife domestication and propagation industry in Zhejiang Province . Journal of Zhejiang A&F University, 2008, 25(1): 109-113.
[15]	CAOMing, LI Wei, ZHOUWei, ZHANG Xing-yong, ZHANG Ren-gong. Foraging sites during the early breeding stage of Syrmaticus humiae in the Nanhua Part of Ailaoshan National Nature Reserve . Journal of Zhejiang A&F University, 2007, 24(2): 203-208.
[16]	HUZhong-jun, WANG Yu, XUEWen-jie, JIANG Hai-rui, XUHong-fa. Population density of Moschus berezovskii in Zibaishan Nature Reserve of Feng County, Shaanxi . Journal of Zhejiang A&F University, 2007, 24(1): 65-71.
[17]	ZHANG Zong, LIANG Nan-nan, GUO Yu-dong, ZHAO Jun. Diversity of landscape architecture approach in city birds protection . Journal of Zhejiang A&F University, 2007, 24(4): 511-515.
[18]	PANShi-xiu, MEN Xiu-xiang, FENG Jin-chao, ZHOUYi-jun, XUHong-fa. A review of studies on habitat selection by small and solitary forest ruminants . Journal of Zhejiang A&F University, 2007, 24(3): 357-362.
[19]	YANG Qiong-xia, YIN Shu, SHENTU Yin, GAO Xin, SUN Yi, HU Jun-xiang. Radioprotective effects of extract from Carya cathayensis nut exocarp on mice . Journal of Zhejiang A&F University, 2006, 23(6): 604-607.
[20]	YAN Shao-jun, ZHU Xi, YU Yi-wu. Construction techniques for the city heronries in Central China . Journal of Zhejiang A&F University, 2006, 23(6): 697-700.

References
Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Get Citation

PDF

XML

Article views(2537) PDF downloads(722) Cited by()

Proportional views

A real-time PCR method for detection of porcine circovirus 2

1. College of Animal Science and Technology, Zhejiang A & F University, Lin’an 311300, Zhejiang, China

Funds:

Received Date: 2015-04-19

Rev Recd Date: 2015-04-19

Publish Date: 2016-04-20

Key words:

Abstract: To develop an accurate and rapid detection method for disease screening and prevention with porcine circovirus type 2 （PCV2）, a single-strand DNA virus that infects pigs, primers targeted to the ORF2 fragment of the PCV2 conserved region were designed. A real time polymerase chain reaction （PCR） method was developed using SYBR Green as a fluorescent dye and serial dilutions of ORF2 recombinant plasmid to construct a standard curve for an absolute quantification. Results showed that the detection limit was obtained with 101 copy DNA per microliter. This method exhibited 1 000 times higher sensitivity than conventional PCR, only specific identification of PCV2, and better repeatability with the less than 2% variation coefficient of intra- or inter-assay experiments. A total of 34 PCV2 positive clinical samples were confirmed using this real time PCR method, demonstrating 100% agreement in comparison with the conventional PCR having only 50% agreement. This real time PCR method could provide a valuable tool for PCV2 prevention and control.［Ch, 4 fig. 4 tab. 26 ref.］

A real-time PCR method for detection of porcine circovirus 2

doi: 10.11833/j.issn.2095-0756.2016.02.023

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Related

Proportional views