Development of anexpress-method for influence and genotyping of H1N1 and H7N9 virus avian influenza a strains by PCR-RFLP analysis

Authors

  • Semen Buriachenko National Science Center Institute of Experimental and Clinical Veterinary Medicine NAAS of Ukraine Pushkinska str., 83, Kharkiv, Ukraine, 61023, Ukraine https://orcid.org/0000-0002-3515-1621
  • Borys Stegniy National Science Center Institute of Experimental and Clinical Veterinary Medicine NAAS of Ukraine Pushkinska str., 83, Kharkiv, Ukraine, 61023, Ukraine https://orcid.org/0000-0002-0661-5837

DOI:

https://doi.org/10.15587/2519-8025.2019.179191

Keywords:

highly pathogenic avianinfluenza A virus H1N1, H7N9, express – diagnostic method for PCR-RFLP, conservative motifs, variable loci, polymorphism

Abstract

Epizootic monitoring in recent years suggests that the highly pathogenic avian influenza A virus (H1N1) and (H7N9) actively circulate in the Eurasian countries. By 2016 - 2019 1.6 thousand outbreaks were recorded. For 2016 - 2019, 1.6 thousand cases of outbreaks were recorded. Of these, there are 872 cases in Europe. The monitoring of infected birds, both migratory and poultry, in places of cross-contact in Ukraine is relevant for preventing outbreaks of epizooties.

The aim of the study. To develop an express method for the identification and determination of bird flu virus A H1N1 and H7N9 strains, based on a polymerase chain reaction with analysis of restriction fragment length polymorphism (PCR-RFLP) of the virus RNA.

Results and discussion. The in silico analysis of the HA, NA, and NP gene amplicons allowed in silico to calculate the primers to the variable loci of the investigated genes, to calculate the reaction conditions, to determine restriction sites for the restriction enzyme to obtain theoretical PCR electrophoregrams. An express method for the detection and identification of influenza A H1N1 and H7N9 virus by three genes (HA, NA, and NP) of H1N1 and H7N9 RNA in polymerase chain reaction, combined with RFLP analysis, was developed. The method of rapid diagnostics is able to detect avian influenza virus A H1N1 and H7N9 and differentiate it from samples of other pathogens of viral infections of birds and animals. It was established, that the PCR-RFLP rapid diagnostic method is able to detect influenza A virus RNA of H1N1 and H7N9 strains with high sensitivity (100 % sensitivity).

Conclusions.The developed method of PCR-based rapid identification, combined with RFLP analysis, makes it possible to significantly simplify the method of identification due to specific amplification of an RNA region having a polymorphic restriction site. Testing of this locus is possible by pre-PCR and restriction of the amplified fragment. The method of express - diagnosis of PLR-RFLP has been established for detecting RNA virus influenza A of high pathogenic H1N1 and H7N9 strains with high indicators of sensitivity (100 % sensitivity)

Author Biographies

Semen Buriachenko, National Science Center Institute of Experimental and Clinical Veterinary Medicine NAAS of Ukraine Pushkinska str., 83, Kharkiv, Ukraine, 61023

Postgraduate student

Department of Infectious Avian Disease

Borys Stegniy, National Science Center Institute of Experimental and Clinical Veterinary Medicine NAAS of Ukraine Pushkinska str., 83, Kharkiv, Ukraine, 61023

Doctor of Veterinary Science, Professor, Academician of the National Academy of Agrarian Sciences of Ukraine, Head of Laboratory

Laboratory for the Study of Viral Poultry Diseases

References

  1. Chen, Y., Liang, W., Yang, S., Wu, N., Gao, H., Sheng, J. et. al. (2013). Human infections with the emerging avian influenza A H7N9 virus from wet market poultry: clinical analysis and characterisation of viral genome. The Lancet, 381 (9881), 1916–1925. doi: http://doi.org/10.1016/s0140-6736(13)60903-4
  2. Gao, R., Cao, B., Hu, Y., Feng, Z., Wang, D., Hu, W. et. al. (2013). Human Infection with a Novel Avian-Origin Influenza A (H7N9) Virus. New England Journal of Medicine, 368 (20), 1888–1897. doi: http://doi.org/10.1056/nejmoa1304459
  3. Neumann, G., Macken, C. A., Kawaoka, Y. (2014). Identification of Amino Acid Changes That May Have Been Critical for the Genesis of A(H7N9) Influenza Viruses. Journal of Virology, 88 (9), 4877–4896. doi: http://doi.org/10.1128/jvi.00107-14
  4. Kageyama, T., Fujisaki, S., Takashita, E., Xu, H., Yamada, S., Uchida, Y. et. al. (2013). Genetic analysis of novel avian A(H7N9) influenza viruses isolated from patients in China, February to April 2013. Euro Surveill, 18 (16), 20453.
  5. Zhang, Q., Shi, J., Deng, G., Guo, J., Zeng, X., He, X. et. al. (2013). H7N9 Influenza Viruses Are Transmissible in Ferrets by Respiratory Droplet. Science, 341 (6144), 410–414. doi: http://doi.org/10.1126/science.1240532
  6. Liu, C. Y., Ai, J. H. (2013). Virological characteristics of avian influenza A H7N9 virus. Zhongguo Dang Dai ErKeZaZhi, 15, 405–408.
  7. Mei, Z., Lu, S., Wu, X., Shao, L., Hui, Y., Wang, J. et. al. (2013). Avian Influenza A(H7N9) Virus Infections, Shanghai, China. Emerging Infectious Diseases, 19 (7), 1179–1181. doi: http://doi.org/10.3201/eid1907.130523
  8. Lv, H., Han, J., Zhang, P., Lu, Y., Wen, D., Cai, J. et. al. (2013). Mild Illness in Avian Influenza A(H7N9) Virus–Infected Poultry Worker, Huzhou, China, April 2013. Emerging Infectious Diseases, 19 (11), 1885–1888. doi: http://doi.org/10.3201/eid1911.130717
  9. Zhou, L., Tan, Y., Kang, M., Liu, F., Ren, R., Wang, Y. et. al. (2017). Preliminary Epidemiology of Human Infections with Highly Pathogenic Avian Influenza A(H7N9) Virus, China, 2017. Emerging Infectious Diseases, 23 (8), 1355–1359. doi: http://doi.org/10.3201/eid2308.170640
  10. Wang, D., Yang, L., Zhu, W., Zhang, Y., Zou, S., Bo, H. et. al. (2016). Two Outbreak Sources of Influenza A (H7N9) Viruses Have Been Established in China. Journal of Virology, 90 (12), 5561–5573. doi: http://doi.org/10.1128/jvi.03173-15
  11. Chang, Y.-F., Wang, W.-H., Hong, Y.-W., Yuan, R.-Y., Chen, K.-H., Huang, Y.-W. et. al. (2018). Simple Strategy for Rapid and Sensitive Detection of Avian Influenza A H7N9 Virus Based on Intensity-Modulated SPR Biosensor and New Generated Antibody. Analytical Chemistry, 90 (3), 1861–1869. doi: http://doi.org/10.1021/acs.analchem.7b03934
  12. Cheng, J., Wang, B., Jiang, X. (2014). Laboratory diagnosis of avian influenza virus H7N9 infection in a renal transplant recipient. International Journal of Clinical and Experimental Medicine, 7, 451–455.
  13. Bouvier, N. M., Lowen, A. C. (2010). Animal Models for Influenza Virus Pathogenesis and Transmission. Viruses, 2 (8), 1530–1563. doi: http://doi.org/10.3390/v20801530
  14. Wang, W., Peng, H., Tao, Q., Zhao, X., Tang, H., Tang, Z. et. al. (2014). Serologic assay for avian-origin influenza A (H7N9) virus in adults of Shanghai, Guangzhou and Yunnan, China. Journal of Clinical Virology, 60 (3), 305–308. doi: http://doi.org/10.1016/j.jcv.2014.04.006
  15. Kageyama, T., Fujisaki, S., Takashita, E., Xu, H., Yamada, S., Uchida, Y. et. al. (2013). Genetic analysis of novel avian A(H7N9) influenza viruses isolated from patients in China, February to April 2013. Eurosurveillance, 18 (15). Available at: https://www.eurosurveillance.org/content/10.2807/ese.18.15.20453-en
  16. Garten, R., Davis, C., Russel, C. et. al. (2009). Antigen and genetic characteristics of swine-origin2009 A(H1N1) influenza virus circulating in humans. Science, 325 (5937), 197–201.
  17. Yoon, J., Yun, S. G., Nam, J., Choi, S.-H., Lim, C. S. (2017). The use of saliva specimens for detection of influenza A and B viruses by rapid influenza diagnostic tests. Journal of Virological Methods, 243, 15–19. doi: http://doi.org/10.1016/j.jviromet.2017.01.013
  18. Lee, M.-S., Chang, P.-C., Shien, J.-H., Cheng, M.-C., Shieh, H. K. (2001). Identification and subtyping of avian influenza viruses by reverse transcription-PCR. Journal of Virological Methods, 97 (1-2), 13–22. doi: http://doi.org/10.1016/s0166-0934(01)00301-9
  19. Radecka, H., Radecki, J. (2016). Electrochemical Sensors for Detections of Influenza Viruses: Fundamentals and Applications. Steps Forwards in Diagnosing and Controlling Influenza. London. doi: http://doi.org/10.5772/64423
  20. Wu, D., Zhang, J., Xu, F., Wen, X., Li, P., Zhang, X. et. al. (2017). A paper-based microfluidic Dot-ELISA system with smartphone for the detection of influenza A. Microfluidics and Nanofluidics, 21 (3). doi: http://doi.org/10.1007/s10404-017-1879-6
  21. Cheng, C., Cui, H., Wu, J., Eda, S. (2017). A PCR-free point-of-care capacitive immunoassay for influenza A virus. Microchimica Acta, 184 (6), 1649–1657. doi: http://doi.org/10.1007/s00604-017-2140-4
  22. Ge, Y., Zhou, Q., Zhao, K., Chi, Y., Liu, B., Min, X. et. al. (2017). Detection of influenza viruses by coupling multiplex reverse-transcription loop-mediated isothermal amplification with cascade invasive reaction using nanoparticles as a sensor. International Journal of Nanomedicine, 12, 2645–2656. doi: http://doi.org/10.2147/ijn.s132670
  23. Karn-orachai, K., Sakamoto, K., Laocharoensuk, R., Bamrungsap, S., Songsivilai, S., Dharakul, T., Miki, K. (2016). Extrinsic surface-enhanced Raman scattering detection of influenza A virus enhanced by two-dimensional gold@silver core–shell nanoparticle arrays. RSC Advances, 6 (100), 97791–97799. doi: http://doi.org/10.1039/c6ra17143e

Downloads

Published

2019-10-11

How to Cite

Buriachenko, S., & Stegniy, B. (2019). Development of anexpress-method for influence and genotyping of H1N1 and H7N9 virus avian influenza a strains by PCR-RFLP analysis. ScienceRise: Biological Science, (3 (18), 9–20. https://doi.org/10.15587/2519-8025.2019.179191

Issue

No. 3 (18) (2019)

Section

Biological Sciences

License

Copyright (c) 2019 Semen Buriachenko, Borys Stegniy

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Our journal abides by the Creative Commons CC BY copyright rights and permissions for open access journals.

Authors, who are published in this journal, agree to the following conditions:

1. The authors reserve the right to authorship of the work and pass the first publication right of this work to the journal under the terms of a Creative Commons CC BY, which allows others to freely distribute the published research with the obligatory reference to the authors of the original work and the first publication of the work in this journal.

 2. The authors have the right to conclude separate supplement agreements that relate to non-exclusive work distribution in the form in which it has been published by the journal (for example, to upload the work to the online storage of the journal or publish it as part of a monograph), provided that the reference to the first publication of the work in this journal is included.