Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Development of a cross-priming isothermal amplification assay based on the glycoprotein B gene for instant and rapid detection of feline herpesvirus type 1


A cross-priming isothermal amplification (CPA) assay was developed for detection of feline herpesvirus type 1 (FHV-1). In this assay, the target fragment of the FHV-1 glycoprotein B gene is amplified rapidly by Bst DNA polymerase at a constant temperature (63 °C, 45 min), using a simple thermostat. The assay had no cross-reactions with four types of feline viruses, and the detection limit was 100 copies/μl. The positive rate of clinical samples from CPA was 100% consistent with qPCR but higher than ordinary PCR, indicating its superiority to ordinary PCR. Visualization was achieved using SYBR Green I dye.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2


  1. 1.

    Sykes JE (2014) Pediatric feline upper respiratory disease. Vet Clin N Am Small Anim Pract 44:331–342. https://doi.org/10.1016/j.cvsm.2013.10.005

  2. 2.

    Maggs DJ (2005) Update on pathogenesis, diagnosis, and treatment of feline herpesvirus type 1. Clin Tech Small Anim Pract 20:94–101. https://doi.org/10.1053/j.ctsap.2004.12.013

  3. 3.

    Litster A, Wu CC, Leutenegger CM (2015) Detection of feline upper respiratory tract disease pathogens using a commercially available real-time PCR test. Vet J 206:149–153. https://doi.org/10.1016/j.tvjl.2015.08.001

  4. 4.

    Tan Y, Dong G, Niu J, Guo Y, Yi S, Sun M, Wang K, Hu G (2019) Development of an indirect ELISA based on glycoprotein B gene for detecting of Feline herpesvirus type 1. Pol J Vet Sci 22:631–633. https://doi.org/10.24425/pjvs.2019.129971

  5. 5.

    Van Ness J, Van Ness LK, Galas DJ (2003) Isothermal reactions for the amplification of oligonucleotides. Proc Natl Acad Sci USA 100:4504–4509. https://doi.org/10.1073/pnas.0730811100

  6. 6.

    Wang HQ, Wu Z, Zhang Y, Tang LJ, Yu RQ, Jiang JH (2012) Label-free genotyping of cytochrome P450 2D6*10 using ligation-mediated strand displacement amplification with DNAzyme-based chemiluminescence detection. Anal Chim Acta 710:111–117. https://doi.org/10.1016/j.aca.2011.10.052

  7. 7.

    Wang H, Kong F, Sorrell TC et al (2009) Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing. BMC Microbiol 9:167. https://doi.org/10.1186/1471-2180-9-167

  8. 8.

    Hira SM, Aledealat K, Chen KS et al (2012) Detection of target ssDNA using a microfabricated Hall magnetometer with correlated optical readout. J Biomed Biotechnol 2012:492730. https://doi.org/10.1155/2012/492730

  9. 9.

    Wozniakowski G, Fraczyk M, Mazur N (2018) Comparison of loop-mediated isothermal amplification (LAMP) and cross-priming amplification (CPA) for detection of African swine fever virus. Pol J Vet Sci 21:827–830. https://doi.org/10.24425/pjvs.2018.125597

  10. 10.

    De Koninck AS, Cnops L, Hofmans M, Jacobs J, Van den Bossche D, Philippe J (2017) Diagnostic performance of the loop-mediated isothermal amplification (LAMP) based illumigene((R)) malaria assay in a non-endemic region. Malar J 16:418. https://doi.org/10.1186/s12936-017-2065-8

  11. 11.

    Gao H, Lei Z, Jia J, Wang S, Chen Y, Sun M, Liang C (2009) Application of loop-mediated isothermal amplification for detection of Yersinia enterocolitica in pork meat. J Microbiol Methods 77:198–201. https://doi.org/10.1016/j.mimet.2009.02.001

  12. 12.

    Babiuk LA, van Drunen LDHS, Tikoo SK (1996) Immunology of bovine herpesvirus 1 infection. Vet Microbiol 53:31–42

  13. 13.

    Ma X, Cui Y, Qiu Z, Zhang B, Cui S (2013) A nanoparticle-assisted PCR assay to improve the sensitivity for rapid detection and differentiation of wild-type pseudorabies virus and gene-deleted vaccine strains. J Virol Methods 193:374–378. https://doi.org/10.1016/j.jviromet.2013.07.018

  14. 14.

    Cao S, Chen H, Zhao J et al (2005) Detection of porcine circovirus type 2, porcine parvovirus and porcine pseudorabies virus from pigs with postweaning multisystemic wasting syndrome by multiplex PCR. Vet Res Commun 29:263–269. https://doi.org/10.1023/b:verc.0000047501.78615.0b

  15. 15.

    Sykes JE, Allen JL, Studdert VP, Browning GF (2001) Detection of feline calicivirus, feline herpesvirus 1 and Chlamydia psittaci mucosal swabs by multiplex RT-PCR/PCR. Vet Microbiol 81:95–108. https://doi.org/10.1016/s0378-1135(01)00340-6

  16. 16.

    Rampazzo A, Appino S, Pregel P, Tarducci A, Zini E, Biolatti B (2003) Prevalence of Chlamydophila felis and feline herpesvirus 1 in cats with conjunctivitis in northern Italy. J Vet Intern Med 17:799–807. https://doi.org/10.1111/j.1939-1676.2003.tb02517.x

  17. 17.

    Binns SH, Dawson S, Speakman AJ, Cuevas LE, Hart CA, Gaskell CJ, Morgan KL, Gaskell RM (2000) A study of feline upper respiratory tract disease with reference to prevalence and risk factors for infection with feline calicivirus and feline herpesvirus. J Feline Med Surg 2:123–133. https://doi.org/10.1053/jfms.2000.0084

  18. 18.

    Maeda K, Kawaguchi Y, Ono M, Tajima T, Mikami T (1995) Restriction endonuclease analysis of field isolates of feline herpesvirus type 1 and identification of heterogeneous regions. J Clin Microbiol 33:217–221

  19. 19.

    Schulz C, Hartmann K, Mueller RS, Helps C, Schulz BS (2015) Sampling sites for detection of feline herpesvirus-1, feline calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease. J Feline Med Surg 17:1012–1019. https://doi.org/10.1177/1098612X15569615

  20. 20.

    Berger A, Willi B, Meli ML, Boretti FS, Hartnack S, Dreyfus A, Lutz H, Hofmann-Lehmann R (2015) Feline calicivirus and other respiratory pathogens in cats with Feline calicivirus-related symptoms and in clinically healthy cats in Switzerland. BMC Vet Res 11:282. https://doi.org/10.1186/s12917-015-0595-2

  21. 21.

    McManus CM, Levy JK, Andersen LA, McGorray SP, Leutenegger CM, Gray LK, Hilligas J, Tucker SJ (2014) Prevalence of upper respiratory pathogens in four management models for unowned cats in the Southeast United States. Vet J 201:196–201. https://doi.org/10.1016/j.tvjl.2014.05.015

  22. 22.

    Fernandez M, Manzanilla EG, Lloret A, Leon M, Thibault JC (2017) Prevalence of feline herpesvirus-1, feline calicivirus, Chlamydophila felis and Mycoplasma felis DNA and associated risk factors in cats in Spain with upper respiratory tract disease, conjunctivitis and/or gingivostomatitis. J Feline Med Surg 19:461–469. https://doi.org/10.1177/1098612X16634387

  23. 23.

    Polak KC, Levy JK, Crawford PC, Leutenegger CM, Moriello KA (2014) Infectious diseases in large-scale cat hoarding investigations. Vet J 201:189–195. https://doi.org/10.1016/j.tvjl.2014.05.020

  24. 24.

    Monne RJ, Kohler K, Kipar A (2018) Calicivirus co-infections in herpesvirus pneumonia in kittens. Vet J 236:1–3. https://doi.org/10.1016/j.tvjl.2018.04.004

  25. 25.

    Nguyen D, Barrs VR, Kelman M, Ward MP (2019) Feline upper respiratory tract infection and disease in Australia. J Feline Med Surg 21:973–978. https://doi.org/10.1177/1098612X18813248

Download references


This work funded was by the National Key Research and Development Program of China (grant number 2016YFD0501002).

Author information

  1. Yuxin Tan and Guoying Dong contributed equally to this work.

    Correspondence to Guixue Hu.

    Ethics declarations

    Ethical approval and consent to participate

    All samples were collected in accordance with the guidelines and regulations of the Animal Care and Use Committee of Jilin Agricultural University, Jilin Province, China. The owners consented to the collection of conjunctival and nasal discharge from the cats.

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Additional information

    Publisher's Note

    Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

    Handling Editor: Tim Skern.

    Rights and permissions

    Reprints and Permissions

    About this article

    Verify currency and authenticity via CrossMark

    Cite this article

    Tan, Y., Dong, G., Xu, H. et al. Development of a cross-priming isothermal amplification assay based on the glycoprotein B gene for instant and rapid detection of feline herpesvirus type 1. Arch Virol 165, 743–747 (2020). https://doi.org/10.1007/s00705-020-04526-5

    Download citation