Establishment and Application of SYBR Green Fluorescence Quantitative PCR for Detection of Porcine Circovirus Type 2

doi:10.7668/hbnxb.2014.03.014

Abstract

Abstract: By quantitative PCR optimization of reaction conditions, a real-time detection of PCV2 SYBR Green quantitative PCR method was established.The results indicated that the method was specificity, the porcine circovirus type 1(PCV1), swine influenza virus(SIV), porcine reproductive and respiratory syndrome virus(PRRSV), pig pseudorabies virus(PRV) and other common diseases of the original pig detection results were negative.The detection limit of the assay was 10 copies/μL of plasmid DNA, 1 000 times higher than that of the routine PCR.The standard curve displayed a linear range from 6.84×10² to 6.84×10⁷ copies and a good reproducibility.

Key words: Porcine circovirus type 2, SYBR Green, Fluorescent quantitative PCR, Standard curve

CLC Number:

S858.28

GUO Hui-juan, LI Xiu-li, ZHANG Guo-wei, YAN Ming-hua, WANG Li-li, ZHANG Li, SUN Ying-feng. Establishment and Application of SYBR Green Fluorescence Quantitative PCR for Detection of Porcine Circovirus Type 2[J]. journal1, 2014, 29(3): 68-73. doi: 10.7668/hbnxb.2014.03.014.

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http://www.hbnxb.net/EN/Y2014/V29/I3/68

References

[1] Guo L J,Lu Y H,Wei Y W,et al.Porcine circovirus type 2(PCV2):Genetic variation and newly emerging genotypes in China [J].Virol J,2010,7:273.
[2] Baylis S A,Finsterbusch T,Bannert N, et a1.Analysis of porcine circovirus type 1 detected in Rotarix vaccine[J].Vaccine,2011,29(4):690-697.
[3] 韩庆功,崔艳红,张智勇.我国猪圆环病毒2型感染的危害[J].黑龙江畜牧兽医,2010(22):84-85.
[4] 田家伟,吴霞,张亮,等.重庆地区某规模化猪场猪圆环病毒2型的PCR检测[J].黑龙江畜牧兽医,2013,3(5):89-90.
[5] 崔尚金,全滟平,李曦,等.猪圆环病毒间接免疫荧光方法的建立[J].中国预防兽医学报,2007,29(1):63-66.
[6] 刘英玉,刘晓丽,廖荣斌,等.利用单抗免疫组化技术检测PCV-2 WH株在人工感染猪组织中的分布[J].中国兽医学报,2010,30(12):1576-1579.
[7] 董志强,张志,李晓成,等.猪圆环病毒PCR 检测方法的优化[J].中国病毒学,2006,21(4):405-407.
[8] 陈小金,陈建飞,时洪艳,等.猪轮状病毒SYBR Green I实时荧光定量PCR检测方法的建立[J].中国兽医科学,2010,40(02):174-179.
[9] 余莹,朱彩霞,杨志彪,等.人双埃克病毒SYBR Green I荧光定量RT-PCR检测方法的建立及初步应用[J].上海交通大学学报:农业科学版,2012,30(1):1-7.
[10] 顾帅,陈陆,常洪涛,等.SYBR Green Ⅰ实时PCR对猪细小病毒体外复制动态分析[J].中国兽医学报,2012,32(3):350-354.
[11] 阳成波,印遇龙,黄瑞林,等.实时定量RT-PCR的原理及方法[J].免疫学杂志,2003,19(S1):145-150.
[12] 来景辉,张堂田,范红结.猪圆环病毒分子生物学研究进展[J].黑龙江畜牧兽医,2013,13(13):24-27.
[13] 张福良,宋长绪,朱道中,等.基于TaqMan探针的猪圆环病毒1型实时荧光定量PCR检测方法的建立[J].中国兽医科学,2007,37(6):477-481.
[14] Turner M J,Medley G F,Woodbine K A,et al.The relationship between porcine circovirus 2 antigen score and antibody titre and histology of lymph nodes in 375 euthanased sick and healthy pigs from 113 British pig farms with and without postweaning multisystemic wasting syndrome[J].Preventive Veterinary Medicine,2009,88(3):213-219.
[15] Brunborg I M,Fossum C,Lium B,et al.Dynamics of serum antibodies to and load of porcine circovirus type 2(PCV2) in pigs in three finishing herds,affected or not by postweaning multisystemic wasting syndrome[J].Acta Veterinaria Scandinavica,2010,52:22.

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