This document describes the biological assay process for testing rabies vaccines. It discusses that the potency of rabies vaccines is determined by comparing the lethal dose of a rabies virus challenge to the dose of vaccine needed to provide protection. It outlines the test animal groups, determination of the challenge virus titre, determination of vaccine potency through serial dilutions, and observation periods. Currently available rabies vaccines in India are also listed.

1. Assay of Rabies
vaccine
M.PHARMACY(PH’CEUTICAL ANALYSIS)
SUBMITTED TO : DR.ALOK SHARMA
SUBMITTED BY : SRI NIWAS
(1st semester)

2. Rabies
 Rabies is a zoonotic disease which is caused by a virus.
 Rabies virus (RABV) belongs to the type species of the genus Lyssavirus
within the family Rhabdoviridae.
 Rabies infects domestic and wild animals, and is spread to people
through close contact with infected saliva through bite, scratch, aerosols
etc.,
 Dogs,
 Bats,
 wild cats,
 Jackals,
 wolves etc

4. RABIES VACCINE
DEFINITION :
Rabies vaccine is a suspension of a suitable strain
of fixed rabies virus grown in suitable approved
cell culture and inactivated by a suitable method.
The vaccine is prepared immediately before use
by reconstitution from the dried vaccine with a
suitable sterile liquid
Dried vaccine + sterile liquid Suspension.

5. BIOLOGICAL ASSAY OF RABIES VACCINE
Principle:
The potency of rabies vaccine is determined by
comparing a lethal intracerebral dose of a rabies
virus with the dose of the standard preparation
of rabies necessary to give for same protection.
Standard preparation: Freeze – dried
preparation.

6. Test animals:
• White mice weight 11gm – 15gm.
• 6 groups of 16 animals , 4 groups
of 10 animals.
• These two groups are used for
titration of LD50 challenge
suspension.
• Injected 0.03ml/mice
intracerebrally.

7. STANDARD CHALLENGE VIRUS SUSPENSION
Standard challenge virus suspension is prepared by injecting
Intracerebrally 0.03ml of 10 fold dilution standard strain horse serum to the test animal
Showing characteristic symptom of encephalitis are sacrified
Harvest the brain aseptically
Wash it with saline solution to remove blood clot
Homogenized brain with 10% digested casein hydrolysate (PH 7.2)
Centrifuge and separated liquid is distributed into sterile ampoules and stored at 2 – 80.

8. DETERMINATION OF CHALLENGE VIRUS
(Determination of virus titre of the challenge virus)
Prepare 3-10 fold dilution of standard challenge virus suspension.
0.03 ml inject intra cerebrally to a groups of 10 mice.
Observe for 14 days
Count the number of mice surviving in each group
Calculate the virus titre of standard challenge virus suspension by
statistical method.

9. DETERMINATION OF POTENCY OF THE VACCINE
Prepare 3-5 fold serial dilution of standard and test solution of vaccines
Separate mice in 6 groups of 16 each
Inject 0.03 ml intra peritoneally and after 7 days prepare same solution and inject
Both standard and test should prepared in such away
After 7 days inject 0.03 ml standard virus suspension to vaccinated mice
Observe if for 14 days and calculate its potency by statistical method.

10. Lowest dilution should protect the
50% of the animal
Highest dilution protect less than
50% of the animal

11. Brand Name Composition Company Packing MRP Rs.
ABHAYRAB inj Inactivated rabies vaccine prepared on vero
cells
HUMAN
BIOLOGICAL
Vial 254.00
RABIPUR inj 2.5 iu inactivated rabies antigen (virus
multiplied in chicken fibroblast cell cultures),
stabilizer (1dose) TEN-haeacel and 0.1%
glutamate q.s.
SANOFI
AVENTIS
Vial 309.00
RABIVAX inj Anti rabies vaccine on human diploid cells SERUM INT. Vial 322.00
VERORAB inj Each freeze dried vaccine contains 1 dose
that the protective activity is > 2.5 iu
RANBAXY Vial 304.00
VEROVAX-R inj Inactivated rabies vaccine prepared on vero
cells
AVENTIS
PASTEUR
0.5ml
prefilled
syringe
281.00
Currently available rabies vaccines in India

12. World Rabies Day is a
cooperative global event
planned to reduce the
suffering from rabies. This
day celebrates Dr. Louis
Pasteur’s vision of a rabies
free world.
WORLD RABIES DAY SEPTEMBER 28

13. REFERENCES
 Wang, H., Zhang, G., Wen, Y ., Yang, S., Xia, X., & Fu, Z. F. (2011). Intracerebral
administration of recombinant rabies virus expressing GM-CSF prevents the
development of rabies after infection with street virus. PloS one, 6(9), e25414
 Hicks, D. J., Fooks, A. R., & Johnson, N. (2012). Developments in rabies vaccines.
Clinical & Experimental Immunology, 169(3), 199-204.
 McGettigan, J. (2010). Experimental rabies vaccines for humans. Expert Rev
Vaccines, 9(10),1177-1186
 Sugiyama, M., & Ito, N. (2007). Control of rabies: epidemiology of rabies in Asia
and development of new-generation vaccines for rabies. Comparative
immunology, microbiology and infectious diseases, 30(5), 273-286.