Preview

Veterinary Science Today

Advanced search

Using spectrometric analysis for indirect estimation of 146S component concentration while measuring FMDV RNA amount

https://doi.org/10.29326/2304-196X-2021-1-36-7-14

Abstract

Foot and mouth disease has a negative impact on economy due to the high cost of eradication campaigns and stringent measures imposed on domestic and international trade in animal products. Prevention and control measures include mass vaccination of susceptible animals and control of post-vaccination immunity level. Concentration of 146S particles, which are the main component affecting the vaccine immunogenicity, is determined during commercial scale production of FMD vaccines. The paper assesses feasibility of spectrometric analysis for indirect determination of 146S component concentration while measuring amount of FMDV RNA isolated after serological binding. This method is cheap, easy-to-use and makes it possible to determine indirectly concentration of FMDV 146S particles in inactivated vaccine raw materials within 3–4 hours. Study of cultural FMDV suspensions shows that the linear model С146S = (3.9 × NRNA 146S + 566,783,689)/ 280,818,944,837 makes it possible to estimate FMDV 146S component concentration in the vaccine raw materials with the help of a spectrometric analysis. The actual results obtained in real-time reverse transcription – polymerase chain reaction (rtRT-PCR) were 97.0–99.9% consistent with the expected results of the spectrometric analysis used to determine cultural FMDV 146S component concentration. When compared to a complement fixation test, the actual results were 94.5–99.5% in line with the expected ones. The actual results for positive control were 99.0–99.6% in line with the expected ones. As expected, no FMDV genome or 146S particles were detected in the negative control sample.

About the Authors

M. I. Doronin
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Maksim I. Doronin, Candidate of Science (Biology), Leading Researcher, Laboratory for FMD Prevention

Vladimir



D. V. Mikhalishin
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Dmitry V. Mikhalishin, Candidate of Science (Veterinary Medicine), Head of Laboratory for FMD Prevention

Vladimir



N. Ye. Kamalova
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Natalia Ye. Kamalova, Doctor of Science (Veterinary Medicine), Chief Researcher, Centre for Preclinical Studies

Vladimir



A. V. Borisov
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
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. 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)

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: 20.02.2020).

4. Martinez-Salas E., Saiz M., Sobrino F. Foot-and-Mouth Disease Virus. In: Animal Viruses: Molecular Biology. Ed. by T. C. Mettenleiter, F. Sobrino. Caister Academic Press; 2008; 1–38.

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. 2016140460/15. Date of filing: 14.10.2016. Date of publication: 18.05.2017. Bull. No. 14. (in Russian)

8. Lozovoy D. A., Mikhalishin D. V., Doronin M. I., Starikov V. A., Guseva M. N., Borisov A. V. Method of spectrometric determination of concentration

9. of 146S foot-and-mouth disease virus particles in non-inactivated raw material for a vaccine by estimating the number of molecules of viral RNA released after immune capture of virions. Patent No. 2712769 Russian Federation, Int. Cl. G01N 33/58 (2006.01), C12Q 1/68 (2006.01). FGBI “ARRIAH”. No. 22019116272. Date of filing: 27.05.2019. Date of publication: 31.01.2020. Bull. No. 4. (in Russian)

10. Chomczynski P., Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat. Protoc. 2006; 1 (2): 581–585. DOI: 10.1038/nprot.2006.83.

11. 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.

12. Dawson R. M. C., Elliott D. C., Elliott W. H., Jones K. M. Data for Biochemical Research. Oxford: Clarendon Press; 1989. 592 р.

13. Glasel J. A. Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. Biotechniques. 1995; 18 (1): 62–63. PMID: 7702855.


Review

For citations:


Doronin M.I., Mikhalishin D.V., Kamalova N.Ye., Borisov A.V. Using spectrometric analysis for indirect estimation of 146S component concentration while measuring FMDV RNA amount. Veterinary Science Today. 2021;1(1):7-14. https://doi.org/10.29326/2304-196X-2021-1-36-7-14

Views: 488


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


ISSN 2304-196X (Print)
ISSN 2658-6959 (Online)