Karimzadeh H, Pakzad Sr, Mahmoudi M, Ajdary S, Norouzi M, Akbari M, Daram M, Jazayeri Jazayeri Sm,
Volume 67, Issue 3 (6-2009)
Background: Hepatitis B
vaccination has been included in routine immunization of all
individuals according to WHO recommendations since 1991. Despite
successful coverage, 3-5% of recipients fail to mount a desirable
protection level of Ab. Vaccine failure results from: emergence of
mutation, immune failure of individuals, decrease in vaccine potency,
and etc. The quality of Hepatitis B vaccine should be evaluated by a
reliable method.
Methods: The amount of vaccine antigen was measured through the in vitro assay
of Hepatitis B vaccines which consists of multiple dilutions of the
reference material and samples. The preparations were evaluated by
Elisa to determine the amount of HBsAg. The data were analyzed by
parallel-line analysis software. The in vivo assay was performed by
inoculating multiple doses of the reference and sample preparations in
Balb/c mice. A control group was also inoculated with vaccine matrix.
Four weeks later, the mice sera were evaluated to determine the
presence of antibodies against Hepatitis B by Elisa method. The data
were analyzed by Probit analysis software.
Results: Both methods were set up in our laboratory by which different batches
of Hepatitis B vaccine were evaluated. It was observed that In vivo and
In vitro methods provide comparable results. Therefore we can use the
in vitro method for routine testing of HB vaccine quality control.
Conclusion: In vitro method can be used in place of In vivo method because of its
time and cost-effectiveness. Moreover, since no animals are used in in
vitro method, it complies well with the 3R concept (Reduction,
Refinement, and Replacement of animal testing) and the current tendency
to use alternative method.
