The Rapid Culture Method for the Indication of Rabies Virus Antigen in Infected Cell Cultures

Current highly sensitive methods for rabies virus and rabies antibodies detection in biological material can be used not only for diagnosis and experimental research, but also for the production of antirabies medicines used for postexposure prophylaxis. The aim of the study was to analyse existing methods for rabies virus and rabies antibodies detection and to assess the potential for using these methods at the control stages during production of heterologous antirabies immunoglobulin obtained from equine serum. The search for cutting-edge highly sensitive in vitro control methods that could compete with the biological method, which is the main method used in antirabies immunoglobulin control, is an important prerequisite for improvement of the production technology and the quality of antirabies medicines. The study demonstrated that the following test methods can be used in the production of antirabies immunoglobulin: fluorescent antibody technique, enzyme-linked immunosorbent assay, cell culture methods, atomic force microscopy, and flow cytometry. These methods could be used alone or as an alternative to the biological method in white mice. These methods were chosen because of their high sensitivity, specificity, rapid and easy implementation, cost-effectiveness, and automatic recording of test results.

show abstract

<i>In vitro</i> methods for rabies virus detection, and evaluation of their use in the production of rabies immunoglobulin

Гаврилова

Генералов

Абрамова

et al. 2021

Biological Products. Prevention, Diagnosis, Treatment

View full text Add to dashboard Cite

show abstract

Development of an in-house reference standard for anti-rabies immunoglobulin potency, to be used in the virus neutralisation test in a Vero cell culture

Абрамова

Gavrilova

Генералов

et al. 2022

Biological Products. Prevention, Diagnosis, Treatment

View full text Add to dashboard Cite

The development and use of new methods of quality control of medicines involve the use of a lot of reference materials in quality control testing. Specialists of the Russian Research Antiplague Institute “Microbe” have proposed an alternative methodological approach to determination of potency of anti-rabies immunoglobulin in cell culture, which requires the development of an in-house reference standard (RS) certified against the biological reference preparation (BRP) of the European Pharmacopoeia¾human rabies immunoglobulin.The aim of the study was to develop and evaluate the metrological characteristics of an in-house RS for anti-rabies immunoglobulin potency, to be used in the virus neutralisation test in a Vero cell culture.Materials and methods: The following materials were used in the study: equine rabies immunoglobulin, Vero continuous cell culture, fixed rabies virus (Moscow 3253Vero strain), human rabies immunoglobulin BRP of the European Pharmacopoeia. The potencies of the candidate in-house RS and rabies immunoglobulin samples were determined in the neutralisation test in cell culture. The results were recorded using a fluorescent microscope. Statistical processing was carried out in accordance with general chapter 1.1.0014.15 of the State Pharmacopoeia of the Russian Federation, 14th edition.Results: the certified value of the in-house RS potency was 180.8±18.8 IU/mL. The confidence limits were determined at the 0.95 probability level. The shelf life of the in-house RS is 1.5 years (when stored according to the sanitary regulation SanPiN 3.3686-21). The certified in-house RS was assigned with the number 41-01-20. A set of technical and operational documentation was developed and approved for the in-house RS. The developed in-house RS can be used for in vitro determination of anti-rabies immunoglobulin potency, expressed in international units, to confirm its compliance with the product specification file.Conclusions: the authors developed an in-house RS for anti-rabies immunoglobulin potency, to be used in the virus neutralisation test in cell culture, certified against the human rabies immunoglobulin BRP of the European Pharmacopoeia.

show abstract

Adaptation of the rabies virus of the production strain "sheep" GNKI to the transplanted cultures of NGUK-1 and ВНК-21/13 CELLS

Galeeva¹,

Ефимова

Arslanova³

et al. 2023

jour

View full text Add to dashboard Cite

The selection of the optimal line of the transplanted cell culture and the adaptation of the rabies virus to it makes it possible to exclude the use of laboratory animals, fully control the process of obtaining viruscontaining material with high infectious activity in large quantities and with a faster and shorter production cycle. The adaptive potential of various strains of rabies virus varies significantly, in this regard, the aim of the study was to study the sensitivity of the production strain of the rabies virus "Sheep" GNKI to the transplanted cultures of NGUK-1 and ВНК-21/13 cells. The adaptation of the rabies virus to the transplanted cultures of NGUK-1 and VNK-21/13 cells was carried out by sequential passivation. The titer of the virus was calculated by the number of fluorescent foci, the concentration of rabies virus antigen was determined by the ELISA method, the pathogenicity of the virus at the level of 11 passages on different cell lines – on white mice.It was found that the transplanted ВНК-21/13 cell line provided a faster adaptation of the virus and the achievement of maximum titers within 36-48 hours, whereas the NGUK-1 line maintained relatively slow replication and ensured the achievement of maximum titers after 96-120 hours. The optimal multiplicity of infection with NGUK-1 and ВНК-21/13 was 0.1 KKID50/cell, while the titers of the "Sheep" virus strains were 4.11±0.26 and 6.17±0.49 lg KKID50/cm3, respectively. Virus replication in ВНК-21/13 cells was characterized by greater intensity: the antigenic titer of the virus at all passage levels was 1.5-2 times (p<0.05) higher than that in NGUK-1 cells. The positive dynamics of the accumulation of the viral titer persisted until passage 8-9, after which this indicator remained stable until passage 11 inclusive. The results of the assessment of the pathogenicity of the rabies virus at the level of passage 11 on different cell lines showed that the virus adapted to both NGUK-1 and ВНК-21/13 has not lost pathogenicity for white mice. Thus, it was found that the transplanted ВНК-21/13 cell line significantly exceeds the NGUK-1 cell line in terms of replicative capabilities and can be used to develop viral raw materials for the production of diagnostic tools and specific prevention.

show abstract

scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.

Contact Info

customersupport@researchsolutions.com

10624 S. Eastern Ave., Ste. A-614

Henderson, NV 89052, USA

This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.

Blog Terms and Conditions API Terms Privacy Policy Contact Cookie Preferences Do Not Sell or Share My Personal Information

Made with 💙 for researchers

Part of the Research Solutions Family.

The Rapid Culture Method for the Indication of Rabies Virus Antigen in Infected Cell Cultures

Cited by 3 publications

References 8 publications

<i>In vitro</i> methods for rabies virus detection, and evaluation of their use in the production of rabies immunoglobulin

<i>In vitro</i> methods for rabies virus detection, and evaluation of their use in the production of rabies immunoglobulin

Development of an in-house reference standard for anti-rabies immunoglobulin potency, to be used in the virus neutralisation test in a Vero cell culture

Adaptation of the rabies virus of the production strain "sheep" GNKI to the transplanted cultures of NGUK-1 and ВНК-21/13 CELLS

Contact Info

Product

Resources

About