VALIDATION OF REAL-TIME REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION FOR INDIRECT ASSESSMENT OF FMD VIRUS TITRE

Procedure for validation of the method for indirect assessment of cultural FMD virus titre (T FMDV) in raw materials used for vaccine production with real time reverse transcription polymerase chain reaction (RT-PCRrt) based on determined amplifcation threshold cycle value (Ct) using linear regression model, TFMDV= –0.2956Ct+ 11.4650, is described in the paper. Testing results for 390 samples of cultural FMD virus were analyzed and basic validation characteristics of the proposed method: specifcity, accuracy, precision, limit of detection and limit of quantifcation, application scope and linearity, were defned. Validation results for the method for indirect assessment of FMD virus titre with RT-PCRrt met the acceptance criteria.

1. Interrelation between infectivity and 146S particle concentration in lapinized foot-and-mouth disease virus suspension [Vzaimosvyaz’ infekcionnosti i koncentracii 146S chastic v suspenzii lapinizirovannogo virusa yashchura]. М. V. Kotova, V. L. Uzyumov, О. G. Andreyeva, М. G. Udalykh. Aktual’nye problemy veterinarnoj virusologii: scientific conference abstracts All-Union Foot and Mouth Disease Research Institute. Vladimir, 1983: 46–48 (in Russian).

2. GOST R ISO 5725-2002. Accuracy (trueness and precision) of measurement methods and results [Tochnost’ (pravil’nost’ i precizionnost’) metodov i rezul’tatov izmerenij]. Part 1–6. Put into force on 01.11.02. М.: Standartinform, 2006 (in Russian).

3. RF National Pharmacopeia Part 1. [Gosudarstvennaya farmakopeya RF]. 12th ed. М.: Scientific Centre for Medical Device Examination, 2008: 254–266 (in Russian).

4. Zhiltsova М. V. Biological properties of epidemic type A, O and Asia-1 foot-and-mouth disease virus isolates [Biologicheskie svojstva ehpizooticheskih izolyatov virusa yashchura tipov A, O i Aziya-1]: author’s abstract … for Candidate of Science degree. Vladimir, 2008 (in Russian).

5. Lakin G. F. Biometrics: text book for Biological Higher Educational Institutes [Biometriya: uchebnoe posobie dlya biol. spec. vuzov]. 4th ed., revised and amended. М.: Vyshaya shkola, 1990 (in Russia).

6. Ponomaryov А. P., Uzyumov V. L., Gruzdev K. N. Foot-and-mouth disease virus: its structure, biological, physical and chemical properties [Virus yashchura: struktura, biologicheskie i fziko-himicheskie svojstva]. Vladimir: Foliant. 2006 (in Russian).

7. Manual of GMP. Validation of analytical methods: theory [Praktikum po GMP. Validaciya analiticheskih metodik: teoriya]. P. Nosyrev, М. Nosyreva, Т. Rasskazova, N. Korneyeva. URL: http://www.nedug.ru/news/фармацевтика/2004/2/12/Практикум-по-GMP--Валидация-аналитическихметодик--теория (access date: 12.11.18) (in Russian).

8. Method for determination of foot-and-mouth disease virus infectivity titre in non-inactivated raw materials used for vaccine production with real-time reverse transcription polymerase chain reaction [Sposob opredeleniya titra infekcionnoj aktivnosti virusa yashchura v neinaktivirovannom syr’e dlya vakciny s primeneniem metoda obratnoj transkripcii i polimeraznoj cepnoj reakcii v rezhime real’nogo vremeni]: pat. 2674076 Russian Federation, MPK7 А61К 39/135. D. V. Mikhalishin, М. I. Doronin, V. А. Starikov [et al.]; FGBI “ARRIAH”. No. 2017145889/10; submitted on 25.12.17; published on 04.12.18 (in Russian).

9. Scherbakov А. V., Timina А. М., Perevozchikova N. А. Development of real-time PCR for foot-and-mouth disease virus detection and quantifcation [Razrabotka PCR v real’nom vremeni dlya obnaruzheniya i kolichestvennogo opredeleniya virusa yashchura]. Russian Veterinary Journal. Productive animals. Special issue devoted to the 50th anniversary of the FGBI “ARRIAH”. 2008; September: 26–27 (in Russian).

10. Implementation of a one-step real-time RT-PCR protocol for diagnosis of foot-and-mouth disease. A. E. Shaw, S. M. Reid, K. Ebert [et al.]. J. Virol. Methods. 2007; 143 (1): 81–85; DOI: 10.1016/j.jviromet.2007.02.009.

11. Principles and methods of validation of diagnostic assays for infectious diseases. OIE. Terrestrial Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Mammals, Birds and Bees). Chapter 1.1.1. URL: http://www.oie.int/fileadmin/Home/eng/Health_standards/aahm/current/chapitre_validation_diagnostics_assays.pdf (access data: 02.11.18).

12. Quality Standard and Guidelines for Veterinary Laboratories: Infectious Diseases. 2nd ed. Paris: OIE, 2008.