We developed and optimized a loop-mediated isothermal amplification (LAMP)-based method to detect porcine parvovirus 7 (PPV7). After using three pairs of specific primers to amplify PPV7 isothermally at 62 °C for 40 min, the amplified product was mixed with SYBR Green I, after which the sample turned green. The method detected PPV7 at concentrations as low as 40 copies/μL, and the sensitivity was consistent with that of nested polymerase chain reaction (PCR) analysis, which was tenfold higher than that of conventional PCR. No cross-reactivity occurred with porcine parvovirus 1, porcine circovirus type 3, porcine circovirus type 2, porcine pseudorabies virus, porcine epidemic diarrhea virus, or porcine reproductive and respiratory syndrome virus. Simultaneous analysis of 76 clinical samples was performed using LAMP, conventional PCR, and nested PCR. The results showed that our method is simple, rapid, sensitive, and specific for the rapid diagnosis of PPV7 in pig farms.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Afolabi KO, Iweriebor BC, Obi LC, Okoh AI (2019) Prevalence of porcine parvoviruses in some South African swine herds with background of porcine circovirus type 2 infection. Acta Trop 190:37–44. https://doi.org/10.1016/j.actatropica.2018.10.010
Cadar D, Csagola A, Kiss T, Tuboly T (2013) Capsid protein evolution and comparative phylogeny of novel porcine parvoviruses. Mol Phylogenet Evol 66(1):243–253. https://doi.org/10.1016/j.ympev.2012.09.030
Cui J, Biernacka K, Fan J, Gerber PF, Stadejek T, Opriessnig T (2017) Circulation of porcine parvovirus types 1 through 6 in serum samples obtained from six commercial polish pig farms. Transbound Emerg Dis 64(6):1945–1952. https://doi.org/10.1111/tbed.12593
Li ZL, Cai YJ, Liang GZ, El-Ashram S, Mei MM, Huang WJ, Li XW, Li WF, He C, Huang SJ (2018) Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP). Sci Rep-Uk. https://doi.org/10.1038/s41598-018-32473-4
Meszaros I, Toth R, Olasz F, Tijssen P, Zadori Z (2017) The SAT protein of porcine parvovirus accelerates viral spreading through induction of irreversible endoplasmic reticulum stress. J Virol. https://doi.org/10.1128/JVI.00627-17
Milek D, Wozniak A, Stadejek T (2018) The detection and genetic diversity of novel porcine parvovirus 7 (PPV7) on Polish pig farms. Res Vet Sci 120:28–32. https://doi.org/10.1016/j.rvsc.2018.08.004
Mori Y, Notomi T (2009) Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemother 15(2):62–69. https://doi.org/10.1007/s10156-009-0669-9
Nagamine K, Hase T, Notomi T (2002) Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probe 16(3):223–229. https://doi.org/10.1006/mcpr.2002.0415
Notomi T, Mori Y, Tomita N, Kanda H (2015) Loop-mediated isothermal amplification (LAMP): principle, features, and future prospects. J Microbiol 53(1):1–5. https://doi.org/10.1007/s12275-015-4656-9
Oh WT, Kim RY, Nguyen VG, Chung HC, Park BK (2017) Perspectives on the evolution of porcine parvovirus. Viruses-Basel. https://doi.org/10.3390/v9080196
Opriessnig T, Fenaux M, Yu S, Evans RB, Cavanaugh D, Gallup JM, Pallares FJ, Thacker EL, Lager KM, Meng XJ, Halbur PG (2004) Effect of porcine parvovirus vaccination on the development of PMWS in segregated early weaned pigs coinfected with type 2 porcine circovirus and porcine parvovirus. Vet Microbiol 98(3–4):209–220. https://doi.org/10.1016/j.vetmic.2003.11.006
Ouh IO, Park S, Lee JY, Song JY, Cho IS, Kim HR, Park CK (2018) First detection and genetic characterization of porcine parvovirus 7 from Korean domestic pig farms. J Vet Sci 19(6):855–857. https://doi.org/10.4142/jvs.2018.19.6.855
Palinski RM, Mitra N, Hause BM (2016) Discovery of a novel Parvovirinae virus, porcine parvovirus 7, by metagenomic sequencing of porcine rectal swabs. Virus Genes 52(4):564–567. https://doi.org/10.1007/s11262-016-1322-1
Sun P, Bai CX, Zhang D, Wang J, Yang KK, Cheng BZ, Li YD, Wang Y (2020) SYBR Green-based real-time polymerase chain reaction assay for detection of porcine parvovirus 6 in pigs. Polish J Vet Sci 23(2):197–202. https://doi.org/10.24425/pjvs.2020.132766
Wang W, Cao L, Sun WC, Xin JL, Zheng M, Tian MY, Lu HJ, Jin NY (2019a) Sequence and phylogenetic analysis of novel porcine parvovirus 7 isolates from pigs in Guangxi China. PLoS ONE. https://doi.org/10.1371/journal.pone.0219560
Wang Y, Yang KK, Wang J, Wang XP, Zhao L, Sun P, Li YD (2019b) Detection and molecular characterization of novel porcine parvovirus 7 in Anhui province from Central-Eastern China. Infect Genet Evol 71:31–35. https://doi.org/10.1016/j.meegid.2019.03.004
Xiao CT, Gerber PF, Gimenez-Lirola LG, Halbur PG, Opriessnig T (2013) Characterization of porcine parvovirus type 2 (PPV2) which is highly prevalent in the USA. Vet Microbiol 161(3–4):325–330. https://doi.org/10.1016/j.vetmic.2012.07.038
Xie ZX, Tang Y, Fan Q, Liu JB, Pang YS, Deng XW, Xie ZQ, Peng Y, Xie LJ, Khan MI (2011) Rapid detection of group I avian adenoviruses by a loop-mediated isothermal amplification. Avian Dis 55(4):575–579. https://doi.org/10.1637/9719-031611-Reg.1
Xing XL, Zhou H, Tong L, Chen Y, Sun YK, Wang H, Zhang GH (2018) First identification of porcine parvovirus 7 in China. Adv Virol 163(1):209–213. https://doi.org/10.1007/s00705-017-3585-9
Xu YG, Cui LC, Wang HW, Huo GC, Li SL (2013) Characterization of the capsid protein VP2 gene of a virulent strain NE/09 of porcine parvovirus isolated in China. Res Vet Sci 94(2):219–224. https://doi.org/10.1016/j.rvsc.2012.09.003
Yu HQ, Cai XQ, Lin ZX, Li XL, Yue QY, Li R, Zhu XQ (2015) Rapid and specific detection of porcine parvovirus using real-time PCR and High Resolution Melting (HRM) analysis. Bmc Vet Res. https://doi.org/10.1186/s12917-015-0364-2
Zhang FF, Ye Y, Song DP, Guo NN, Peng Q, Li AQ, Zhou XR, Chen YJ, Zhang M, Huang DY, Tang YX (2017) A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification. Biol Res. https://doi.org/10.1186/s40659-017-0135-6
Zheng S, Wu X, Shi J, Peng Z, Gao M, Xin C, Liu Y, Wang S, Xu S, Han H, Yu J, Sun W, Cong X, Li J, Wang J (2018) Rapid specific and visible detection of porcine circovirus type 3 using loop-mediated isothermal amplification (LAMP). Transbound Emerg Dis 65(3):597–601. https://doi.org/10.1111/tbed.12835
We would like to thank Editage (www.editage.cn) for English language editing.
This work was supported financially by Anhui Provincial Primary Research & Developement Plan ( 202004f06020008, 201904f06020030) and Ningbo Health Branding Subject Fund (No. ppxk2018-10).
Conflict of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
All experiments were compliant with the ethical standards of Anhui Agricultural University.
About this article
Cite this article
Wang, Y., Zhang, D., Bai, C. et al. Using loop-mediated isothermal amplification for visual detection of porcine parvovirus 7. 3 Biotech 11, 111 (2021). https://doi.org/10.1007/s13205-020-02623-5
- Porcine parvovirus 7
- Loop-mediated isothermal amplification
- Rapid detection