Determination of FMDV 146S particle concentration by spectrometric method during viral RNA quantification
https://doi.org/10.29326/2304-196X-2020-4-35-304-312
Abstract
During FMD vaccine production, special attention is paid to the concentration of 146S particles bearing the critical biological features of FMDV and being the main components that have an effect on vaccine immunogenicity. For this reason, each batch of vaccine raw material is tested for 146S component concentration. The paper presents the results of the use of a spectrometric method for whole particle concentration determination during quantification of FMDV RNA extracted after immune capture. It is an inexpensive, easy-to-perform method allowing for determination of FMDV 146S particle concentration in the non-inactivated culture suspension. 146S particle concentration was found to depend on the number of RNA molecules extracted from virions after their strain-specific immune capture and quantitatively detected by the spectrometric method. The presented method allows for determination of 146S component concentration in the non-inactivated vaccine raw material using the proposed linear model. The spectrometric method showed 94.5–99.5% correlation with real time reverse transcription polymerase chain reaction and complement fixation test based on the results of tests of 360 non-inactivated suspensions of FMDV of all types. Tests of the positive control demonstrated 99.0–99.6% compatibility of actual and expected results. FMDV genome and 146S particles were not detected in the negative control, and that was in line with expectations.
About the Authors
M. I. DoroninRussian Federation
Maksim I. Doronin, Candidate of Science (Biology), Senior Researcher, Laboratory for FMD Prevention
Vladimir
D. V. Mikhalishin
Russian Federation
Dmitry V. Mikhalishin, Candidate of Science (Veterinary Medicine), Head of Laboratory for FMD Prevention
Vladimir
V. A. Starikov
Russian Federation
Vyacheslav A. Starikov, Candidate of Science (Veterinary Medicine), Leading Researcher, Laboratory for FMD Prevention
Vladimir
D. A. Lozovoy
Russian Federation
Dmitry A. Lozovoy, Doctor of Science (Veterinary Medicine), Associate Professor
Vladimir
Yu. S. El’kina
Russian Federation
Yulia S. El’kina, Post-Graduate Student, Technologist, Laboratory for FMD Prevention
Vladimir
A. V. Borisov
Russian Federation
Alexey V. Borisov, Candidate of Science (Veterinary Medicine), Leading Researcher, Laboratory for FMD Prevention
Vladimir
References
1. Foot and mouth disease (infection with foot and mouth disease virus). In: OIE. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. 2018; Chap. 3.1.8: 433–464. Available at: https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.01.08_FMD.pdf.
2. Lubroth J., Rodriguez L., Dekker A. Vesicular diseases. In: Diseases of Swine. Ed. by B. E. Straw, J. J. Zimmerman, S. D’Allaire, D. J. Taylor. 9th ed. Ames, Iowa, USA: Blackwell Publishing Professional; 2006: 517–536.
3. Nucleotide Database of National Center for Biotechnology Information (NCBI). Available at: https://www.ncbi.nlm.nih.gov/nuccore/?term=FMDV+complete (date of access: 25.02.2019).
4. Ponomarev А. P., Uzyumov V. L. Foot-and-mouth disease virus: structure, biological, physical and chemical properties [Virus yashchura: struktura, biologicheskie i fiziko-himicheskie svojstva]. Vladimir: Foliant; 2006. 250 p. (in Russian)
5. Alexandersen S., Zhang Z., Donaldson A. L., Garland A. J. M. The pathogenesis and diagnosis of foot-and-mouth disease. J. Comp. Pathol. 2003; 129 (1): 1–36. DOI: 10.1016/s0021-9975(03)00041-0.
6. Bondarenko А. F. Qualitative and quantitative immunochemical assay of viral proteins [Kachestvennyj i kolichestvennyj immunohimicheskij analiz virusnyh belkov]. Suzdal; 1994. 92 p. (in Russian)
7. Lozovoy D. A., Mikhalishin D. V., Doronin M. I., Shcherbakov A. V., Timina A. M., Shishkova A. A., et al. Method for foot and mouth disease virus 146S-component concentration determination in virus-containing raw material for vaccine using reverse transcription-polymerase chain reaction method in real time mode. Patent No. 2619878 Russian Federation, Int. Cl. G01N 33/58 (2006.01), C12Q 1/68 (2006.01). FGBI “ARRIAH”. No. 2016140460. Date of filing: 14.10.2016. Date of publication: 18.05.2017. Bull. No. 14. (in Russian)
8. Chomczynski P., Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twentysomething years on. Nat. Protoc. 2006; 1 (2): 581–585. DOI: 10.1038/ nprot.2006.83.
9. Peirson S. N., Butler J. N. RNA extraction from mammalian tissues. In: Circadian Rhythms. Methods in Molecular Biology™. Ed. by E. Rosato. 2007; 362: 315–327. DOI: 10.1007/978-1-59745-257-1_22.
10. Vladimirov Yu. A., Potapenko A. Ya. Physics and chemistry of photobiological processes [Fiziko-himicheskie osnovy fotobiologicheskih processov]. М.: Vysshaya shkola; 1989; 20–22. (in Russian)
11. Kaporsky L. N. Optical density [Opticheskaya plotnost’]. In: Encyclopedia of Physics. Vol. 3: Magnetoplasmic – Poynting’s theorem [Fizicheskaya enciklopediya. T. 3: Magnitoplazmennyj – Pojntinga teorema]. Editor-in-Chief A. M. Prokhorov. M.: Great Soviet Encyclopedia;1992: 441. (in Russian)
12. Glasel J. A. Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. Biotechniques. 1995; 18 (1): 62–63. PMID: 7702855.
13. Dawson R. M. C., Elliott D. C., Elliott W. H., Jones K. M. Data for Biochemical Research. Oxford: Clarendon Press; 1989. 592 р.
14. Newell D. B. A more fundamental International System of Units. Physics Today. 2014; 67 (7): 35–41. DOI: 10.1063/PT.3.2448.
15. Strohmaier K., Adam K.-H. Die Struktur des Virus der Maulund Klauenseuche. Zentralblatt für Veterinärmedizin. Reihe B. 1976; 23 (5/6): 483– 506. DOI: 10.1111/j.1439-0450.1976.tb01628.x
Review
For citations:
Doronin M.I., Mikhalishin D.V., Starikov V.A., Lozovoy D.A., El’kina Yu.S., Borisov A.V. Determination of FMDV 146S particle concentration by spectrometric method during viral RNA quantification. Veterinary Science Today. 2020;(4):304-312. https://doi.org/10.29326/2304-196X-2020-4-35-304-312