Validation and Standardization of Virus Neutralizing Test Using Indirect Immunoperoxidase Technique for the Quantification of Antibodies to Rabies Virus

Abstract

A virus neutralizing test using an indirect immunoperoxidase technique (VNT-IIP) for rabies has been developed for the titration of dog and cat serum samples in Japan. The VNT-IIP has the advantage that results obtained can be viewed by the naked eye. The purpose of this study was to validate the VNT-IIP and compare it with one of the international standard methods, the fluorescent antibody virus neutralization test (FAVNT). The VNT-IIP showed satisfactory repeatability, high analytical specificity and good accuracy. Regarding the comparison between the VNT-IIP and the FAVNT, the VNT-IIP showed good agreement (91.9%), high sensitivity (92.8%) as well as specificity (87.0%) and good correlation (r = 0.92). As described above, the validation of the VNT-IIP was satisfactory and the performances of the test proved to be equivalent to those of an international standard method.

Inhibition ofPasteurella multocidaAdhesion to Rabbit Respiratory Epithelium Using Lectins

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Development and evaluation of a new immunohistochemistry-based test for the detection of rabies virus neutralizing antibodies

“…Two previous studies have reported the utility of an indirect immunoperoxidase test using polyclonal anti-rabies antisera for the detection of RVNA in serum samples. 10,11 In the present study, the sensitivity and specificity of the test were enhanced further with the use of a biotinylated anti-N monoclonal antibody cocktail, which was previously shown to be 100% specific and sensitive when employed in detection of rabies virus N antigen in brain smears in our laboratory.…”

“…11 Serial dilutions of the test sera was added to each serum dilution. The plates were incubated at 37 °C in a CO 2 incubator (NuAire) for one hour.…”

Rabies Immune Status of Dogs Brought into the Hyogo Prefecture Animal Well-Being Center, Japan

“…The remaining 60 serum samples were from dogs captured and brought into the Center by rabies prevention officers or others in 2007. The neutralizing antibody titers for rabies were measured using the indirect immunoperoxidase virus neutralizing test [5]. In brief, a cell suspension of Neuro-2a (ATCC CCL-131) was prepared in 96-well plates.…”