- 中文核心期刊
- CSCD来源期刊
- 中国科技核心期刊
- CA、CABI、ZR收录期刊
| Citation: |
LIN Q，KANG L B，WU R S，et al. A TaqMan RT-qPCR Assay for Detecting Porcine Rotavirus Group A[J]. Fujian Journal of Agricultural Sciences，2025，40（4） ：370−376. DOI: 10.19303/j.issn.1008-0384.2025.04.006
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A rapid method for detecting porcine rotavirus group A (PoRV A) in disease monitoring and epidemiological investigation was developed.
Specific primers and probes were selected based on the VP6 gene sequences of PoRV A in GenBank (accession numbers MT025937.1, OP978242.1 and PP566178.1). Reaction conditions of the TaqMan RT-qPCR method were optimized followed by evaluations on assay specificity, sensitivity, repeatability, and clinical application.
The newly developed method could specifically amplify PoRV nucleic acid with a minimum detection limit of 27.0 copies·μL−1 and a sensitivity 100 times higher than RTPCR. There was no cross reactivity with the nucleic acids of PEDV, TGEV or PDCoV. The method showed high repeatability with the variation coefficients on intra- and inter-group below 1.10%. On 151 clinical specimens suspected of PoRV, a detection rate of 42.38% (64/151), which was better than that of conventional RT-PCR at 33.11% (50/151), was obtained by the assay.
The new TaqMan RT qPCR method for detecting VP6 gene of PoRV A was high in assay sensitivity, specificity, and repeatability. It was considered suitable for PoRV detection and epidemiological investigations.
| [1] |
黄校花. 抗猪轮状病毒VP6单克隆抗体的制备及双抗体夹心ELISA检测方法的建立[D]. 南昌:江西农业大学,2023.
HUANG X H. Preparation of monoclonal antibodies against porcine rotavirus VP6 and establishment of a double antibody sandwich ELISA assay[D]. Nanchang:Jiangxi Agricultural University,2023. (in Chinese)
|
| [2] |
MIYABE F M,DALL AGNOL A M,LEME R A,et al. Porcine rotavirus B as primary causative agent of diarrhea outbreaks in newborn piglets[J]. Scientific Reports,2020,10(1) :22002. DOI: 10.1038/s41598-020-78797-y
|
| [3] |
VLASOVA A N,AMIMO J O,SAIF L J. Porcine rotaviruses:Epidemiology,immune responses and control strategies[J]. Viruses,2017,9(3) :48. DOI: 10.3390/v9030048
|
| [4] |
常新见. 华东地区猪轮状病毒分子流行病学调查及亚单位疫苗初步研究[D]. 拉萨:西藏大学,2020.
CHANG X J. The investigation of molecular epidemiological of porcine rotavirus in East China and subunit vaccine initial research[D]. Lasa:Tibet University,2020. (in Chinese)
|
| [5] |
谢淑敏. 猪轮状病毒病的特征及防控措施[J]. 广东饲料,2023,32(6) :49−51. DOI: 10.3969/j.issn.1005-8613.2023.06.019
XIE S M. Characteristics,prevention and control measures of porcine rotavirus disease[J]. Guangdong Feed,2023,32(6) :49−51. (in Chinese) DOI: 10.3969/j.issn.1005-8613.2023.06.019
|
| [6] |
燕贺,孙玉洁,赵文影,等. 中国不同地区猪轮状病毒的分离鉴定及分子流行病学分析[J]. 中国预防兽医学报,2024,46(2) :201−205. DOI: 10.3969/j.issn.1008-0589.202305008
YAN H,SUN Y J,ZHAO W Y,et al. Isolation and molecular epidemiological analysis of porcine rotavirus in some regions of China[J]. Chinese Journal of Preventive Veterinary Medicine,2024,46(2) :201−205. (in Chinese) DOI: 10.3969/j.issn.1008-0589.202305008
|
| [7] |
樊高,申翰钦,王连想,等. 猪轮状病毒的分离鉴定和全基因组序列分析[J]. 中国兽医杂志,2023,59(6) :47−54.
FAN G,SHEN H Q,WANG L X,et al. Isolation,identification and whole genome sequence analysis of porcine rotavirus[J]. Chinese Journal of Veterinary Medicine,2023,59(6) :47−54. (in Chinese)
|
| [8] |
李元新,陈伯祥,赵子惠,等. 猪轮状病毒病诊断与防控研究进展[J]. 畜牧兽医杂志,2022,41(5) :58−61,64. DOI: 10.3969/j.issn.1004-6704.2022.05.018
LI Y X,CHEN B X,ZHAO Z H,et al. Research progress on the diagnosis and control of porcine rotavirus disease[J]. Journal of Animal Science and Veterinary Medicine,2022,41(5) :58−61,64. (in Chinese) DOI: 10.3969/j.issn.1004-6704.2022.05.018
|
| [9] |
孟柳. 抗猪轮状病毒单克隆抗体的制备及双抗体夹心ELISA检测方法的初步建立[D]. 哈尔滨:东北农业大学,2014.
MENG L. Development of monoclonal antibodies against porcine rotavirus and double antibodies sandwich ELISA for the virus detection[D]. Harbin:Northeast Agricultural University,2014. (in Chinese)
|
| [10] |
潘向英,曾智勇,梁海英,等. 猪轮状病毒VP6蛋白截短表达及间接ELISA抗体检测方法的建立[J]. 中国畜牧兽医,2025,52(3) :1231−1240.
PAN X Y,ZENG Z Y,LIANG H Y,et al. Truncated expression of porcine rotavirus VP6 protein and establishment of an indirect ELISA antibody detection method[J]. China Animal Husbandry & Veterinary Medicine,2025,52(3) :1231−1240. (in Chinese)
|
| [11] |
GHONAIM A H,YI G Y,LEI M K,et al. Isolation,characterization and whole-genome analysis of G9 group a rotaviruses in China:Evidence for possible Porcine–Human interspecies transmission[J]. Virology,2024,597:110129. DOI: 10.1016/j.virol.2024.110129
|
| [12] |
TAO R,CHANG X J,ZHOU J Z,et al. Molecular epidemiological investigation of group A porcine rotavirus in East China[J]. Frontiers in Veterinary Science,2023,10:1138419. DOI: 10.3389/fvets.2023.1138419
|
| [13] |
陈秋勇,陈渊泉,陈如敬,等. PEDV、TGEV双重 Taq Man实时荧光定量PCR方法的建立及病原学调查[J]. 福建农业学报,2022,37(11) :1381−1387.
CHEN Q Y,CHEN Y Q,CHEN R J,et al. Duplex Taqman qPCR for detecting porcine epidemic diarrhea and transmissible gastroenteritis viruses and epidemic study in Fujian[J]. Fujian Journal of Agricultural Sciences,2022,37(11) :1381−1387. (in Chinese)
|
| [14] |
CHEN Q Y,SUN Z H,CHE Y L,et al. High prevalence,genetic diversity,and recombination of Porcine sapelovirus in pig farms in Fujian,Southern China[J]. Viruses,2023,15(8) :1751. DOI: 10.3390/v15081751
|
| [15] |
梁子英,刘芳. 实时荧光定量PCR技术及其应用研究进展[J]. 现代农业科技,2020(6) :1−3,8. DOI: 10.3969/j.issn.1007-5739.2020.06.001
LIANG Z Y,LIU F. Research progress on real-time quantitative PCR technology and its application[J]. Modern Agricultural Science and Technology,2020(6) :1−3,8. (in Chinese) DOI: 10.3969/j.issn.1007-5739.2020.06.001
|
| [16] |
陈小飞,严楠,张斌,等. 猪A群轮状病毒荧光定量PCR检测方法的建立及应用[J]. 动物医学进展,2021,42(2) :41−45. DOI: 10.3969/j.issn.1007-5038.2021.02.007
CHEN X F,YAN N,ZHANG B,et al. Establishment and application of TB green real-time fluorescence quantitative RT-PCR for detection of porcine RVA[J]. Progress in Veterinary Medicine,2021,42(2) :41−45. (in Chinese) DOI: 10.3969/j.issn.1007-5038.2021.02.007
|
| [17] |
汤学超. 猪轮状病毒VP4蛋白截短体的原核表达及免疫原性研究[D]. 荆州:长江大学,2023.
TANG X C. Prokaryotic expression and immunogenicity of truncated VP4 protein of porcine rotavirus[D]. Jingzhou:Yangtze University,2023. (in Chinese)
|
| [18] |
张子微. 湖北地区猪A群轮状病毒的流行病学调查与VP7基因的遗传分析[D]. 荆州:长江大学,2023.
ZHANG Z W. Epidemiological investigation and genetic analysis of VP7 gene of group A swine rotavirus in Hubei province[D]. Jingzhou:Yangtze University,2023. (in Chinese)
|
| [19] |
乔成鹏. 2015~2016年中国部分地区猪轮状病毒感染检测及病毒分离与鉴定[D]. 大庆:黑龙江八一农垦大学,2019.
QIAO C P. Detection,isolation and identification of porcine rotavirus infection in some areas of China from 2015 to 2016[D]. Daqing:Heilongjiang Bayi Agricultural University,2019. (in Chinese)
|
| [20] |
杜庆川. 江苏省规模猪场主要病毒性疾病的流行病学调查[D]. 呼和浩特:内蒙古农业大学,2023.
DU Q C. Epidemiological investigation of major viral diseases in large-scale pig farms in Jiangsu Province[D]. Hohhot:Inner Mongolia Agricultural University,2023. (in Chinese)
|
| [21] |
刘云波,赵洪翠,王志成,等. 猪病毒性腹泻分子流行病学调查[J]. 中国畜牧兽医,2013,40(2) :204−207. DOI: 10.3969/j.issn.1671-7236.2013.02.049
LIU Y B,ZHAO H C,WANG Z C,et al. Molecular epidemiological investigation of swine viral diarrhea[J]. China Animal Husbandry & Veterinary Medicine,2013,40(2) :204−207. (in Chinese) DOI: 10.3969/j.issn.1671-7236.2013.02.049
|
| [22] |
何晓明,田小艳,王东东,等. 2021—2022年我国部分地区猪轮状病毒分子流行病学调查[J]. 畜牧与兽医,2024,56(3) :77−85.
HE X M,TIAN X Y,WANG D D,et al. Molecular epidemiology investigation of porcine rotavirus in some regions of China in 2021-2022[J]. Animal Husbandry & Veterinary Medicine,2024,56(3) :77−85. (in Chinese)
|
| [23] |
谷拉强,陶然,程曦,等. 2022—2023年我国部分地区猪A群轮状病毒分子流行病学调查分析[J]. 南方农业学报,2023,54(10) :3083−3091. DOI: 10.3969/j.issn.2095-1191.2023.10.027
GU L Q,TAO R,CHENG X,et al. Molecular epidemiological survey of group A porcine rotavirus in selected areas of China during 2022-2023[J]. Journal of Southern Agriculture,2023,54(10) :3083−3091. (in Chinese) DOI: 10.3969/j.issn.2095-1191.2023.10.027
|
| [24] |
田红利,柳佳佳,梁海英,等. 贵州省2017年–2023年PEDV和Po RVA流行病学调查及Po RVA VP7、VP4基因的遗传演化分析[J]. 中国预防兽医学报,2024,46(8) :791−798. DOI: 10.3969/j.issn.1008-0589.202312018
TIAN H L,LIU J J,LIANG H Y,et al. Epidemiological investigation on the prevalence of PEDV and Po RVA in Guizhou during 2017–2023 and genetic evolution analysis of VP7 and VP4 genes of Po RVA strains[J]. Chinese Journal of Preventive Veterinary Medicine,2024,46(8) :791−798. (in Chinese) DOI: 10.3969/j.issn.1008-0589.202312018
|
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