猪嵴病毒RT-PCR检测方法的建立及应用

Abstract

Abstract: A detection method for porcine kobuvirus (PKV) with RT-PCR technique was established to detect the 3D gene of PKV from the intestinal tract of diarrhea piglets, with the detection limit about 2.3 pg RNA. 50 samples of diarrhea piglets were detected by the RT-PCR method, the PKV positive rate of samples was 58 percent. The PCR detection method established in this study has the characteristics of sensitivity, high specificity and good repeatability, and could be used in molecular epidemiological investigation and rapid clinical diagnosis of PKV.

Key words: porcine kobuvirus; RT-PCR; homology analysis

CLC Number:

S852.65

ZHANG Wen-bo, LENG Chuang, DUAN Jun, DENG Shun-zhou. Establishment and Application of RT-PCR Detection of Pocrine Kobuvirus[J]. , 2013, 40(1): 42-46.

References

1 An D J, Jeoung H Y, Jeong W,et al.Porcine kobuvirus from pig stool in Korea[J]. Virus Genes,2011,42(2):208~211.
2 Barry A F, Ribeiro J, Alfieri A F,et al. First detection of kobuvirus in farm animals in Brazil and the Netherlands[J].Infect Genet Evol, 2011,11(7):1811~1814.
3 Chen J, Liu X, Shi D,et al. Complete genome sequence of a porcine epidemic diarrhea virus variant[J]. J Virol,2012,86(6):3408.
4 Fan J H, Zuo Y Z, Li J H,et al. Heterogeneity in membrane protein genes of porcine epidemic diarrhea viruses isolated in China[J].Virus Genes,2012,45(1):113~117.
5 Khamrin P, Maneekarn N, Kongkaew A,et al. Porcine kobuvirus in piglets, Thailand[J].Emerg Infect Dis,2009,15(12):2075~2076.
6 Lin Y, Xiao S, Zeng Y,et al. Complete genome sequence of porcine kobuvirus strain WUH1[J].J Virol,2012,86(12):7010.
7 Li Z L, Zhu L, Ma J Y,et al. Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) field strains in south China[J]. Virus Genes, 2012,45(1):181~185.
8 Park S J, Kim H K, Moon H J,et al. Molecular detection of porcine kobuviruses in pigs in Korea and their association with diarrhea[J].Arch Virol,2010,155(11):1803~1811.
9 Park S J, Kim H K, Song D S, et al. Molecular detection and genetic characterization of kobuviruses in fecal samples collected from diarrheic cattle in Korea[J].Infect Genet Evol,2011,11(5):1178~1182.
10 Reuter G, Boldizsár A, Kiss I,et al.Candidate new species of kobuvirus in porcine hosts[J].Emerg Infect Dis,2008,14(12):1968~1970.
11 Reuter G, Boldizsár A, Pankovics P. Complete nucleotide and amino acid sequences and genetic organization of porcine kobuvirus, a member of a new species in the genus Kobuvirus, family Picornaviridae[J].Arch Virol,2009, 154(1):101~108.
12 Reuter G, Boros A, Pankovics P. Kobuviruses——a comprehensive review[J]. Rev Med Virol,2011,21(1):32~41.
13 Song D, Park B. Porcine epidemic diarrhoea virus: A comprehensive review of molecular epidemiology, diagnosis, and vaccines[J]. Virus Genes,2012,44(2):167~175.
14 Wang C, Lan D, Hua X.Porcine kobuvirus from pig stool specimens in Shanghai, China[J]. Virus Genes,2011,43(3):350~352.
15 Wang C, Lan D, Cui L,et al. Molecular characterization of a porcine kobuvirus strain in China[J].Arch Virol,2012,157(3):573~578.
16 Yu J M, Jin M, Zhang Q,et al. Candidate porcine kobuvirus, China[J].Emerg Infect Dis,2009,15(5):823~825.
17 Yu J M, Xu Z Q, Li B W,et al. Analysis and characterization of the complete genome of a member of a new species of kobuvirus associated with swine[J].Arch Virol, 2011, 156(5):747~751.

[1]	ZHANG Xinjie, XUE Shaohua, LYU Zixin, HUANG Xirong, CHEN Yuxuan, FAN Kewei, YANG Xiaoyan, DAI Ailing. Investigation of Co-infection of PPV7 and PCV2 and Genetic Variation Analysis for PPV7 Cap Gene in Fujian and Guangdong [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2291-2297.
[2]	LI Mingyang, FANG Qingli, HUANG Weijian. Molecular Epidemiology of Bovine Astrovirus in Buffalo from Guangxi [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2926-2934.
[3]	YAN Xiukui, HAN Zhixiao, DU Qian, WANG Wei, ZHU Yuanzhi, XIN Jialiang, YI Chizhe, BI Jingshan, SUN Wenchao, ZHENG Min. Detection and Sequence Analysis of Canine Circovirus in Chongqing [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3069-3077.
[4]	LI Xiaofei, CHEN Ting, WEI Xiaoxiao, SUN Airong, HUANG Yan, GE Xiaojie, XU Ting, WANG Caihong, QIN Liting. Establishment and Application of TaqMan-MGB Real-time RT-PCR Assay for Detection of Porcine Reproductive and Respiratory Syndrome Virus [J]. , 2019, 46(1): 239-246.
[5]	YANG Fan, SHI Shunli, XU Na, LEI Yu, CHANG Tana, LI Pingan, GUAN Pingyuan. Establishment of a Multiplex RT-PCR Detection Method for Three Genotypes of Bovine Parainfluenza Virus Type 3 [J]. , 2018, 45(1): 32-38.
[6]	ZHANG Xiao-fen, YE Gui-sheng. PCR Amplification and Bioinformatics Analysis of IL-7 Gene in Tibetan Sheep [J]. , 2017, 44(9): 2558-2565.
[7]	TANG Lian-fei, YU Si-yu, ZHOU Yu. Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype [J]. , 2017, 44(6): 1847-1853.
[8]	KONG Jing-ya, GUO Qiang-qiang, KONG Ke-wei, LI Jie, ZHOU Xia, QI Ya-yin. Detection on Biofilm and Biofilm-associated Genes of Enterococcus faecalis Inducing Lamb Encephalitis [J]. , 2017, 44(3): 679-685.
[9]	SHI Kai-chuang, XU Xin-ting, HU Jie, SU Yan-qiong, LU Wen-jun, CHEN Han-zhong. Establishment and Application of a Multiplex RT-PCR Assay for Differential Detection of Classical, Highly Pathogenic and Vaccine Strains of North American Genotype PRRSV [J]. , 2017, 44(3): 879-887.
[10]	YAO Huai-bing, ZHAO Yi, LIU Hong, ZHANG Yi, REN Fang, HUANG Jiong. Development and Application of Two-temperature RT-PCR for Detectionof Type O Foot and Mouth Disease Virus [J]. , 2017, 44(2): 311-318.
[11]	HUANG Xiao-xing, WANG Xian, JIN Wen-jie. Research Advances in Pathogenicity and Detection Techniques of Porcine Kobuvirus [J]. , 2017, 44(12): 3466-3472.
[12]	WANG Bo, WANG Hui-yu, ZHAO Bao-hua, LUO Jing, WANG Cheng-min, HE Hong-xuan, HAN Xue-qing. Development and Application of Duplex Real-time RT-PCR Assay for Detection of H1 and H3 Subtype Swine Influenza Viruses [J]. , 2017, 44(10): 3035-3041.
[13]	MA Ping, LI Da, TANG De-yuan, ZHANG Yuan-xin, ZHANG Hua, ZENG Zhi-yong, LIU Xia, WEI Guan-dong. Establishment and Application of Single and Duplex RT-PCR Method for Differentiation of Wild-type and Vaccine Viruses of Classical Swine Fever Virus [J]. , 2016, 43(9): 2230-2239.
[14]	HU Hong-hui, NAN Wen-jin, HUANG Jian-qiang, WU Jing-bo, PENG Guo-liang. Establishment and Application of a Double RT-PCR Method for Detection of PEDV and TGEV [J]. , 2016, 43(9): 2279-2284.
[15]	LIU Mei-fen, WANG Shu-juan, YAN Ruo-qian, WANG Dong-fang, CAO Wei-wei, ZHAO Xue-li, LI Qin-nan, LI Ning. Establishment and Application of Nested RT-PCR Assay for Detection of Classical Swine Fever Virus [J]. , 2016, 43(9): 2285-2290.

Establishment and Application of RT-PCR Detection of Pocrine Kobuvirus

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments