Fig 1.
Flowchart showing current and proposed (dashed lines) laboratory testing regimen for diagnosis of rabies in Indonesia following competency assessment of individual provincial laboratories using the RIADacetone test.
Fig 2.
Expression and purification of RABV NP and characterization of antiserum.
RABV NP inclusion bodies (IBs) (A) and gel-purified RABV NP (B) were resolved by SDS PAGE and stained with Coomassie blue. All lanes of gels stained with Coomassie blue were loaded with 10 μl of RABV NP in the dilutions or amounts indicated. Recombinant, gel-eluted His-tagged RABV NP was identified by immunoblotting with anti-His antibody (1:1,000) followed by sheep anti-mouse-HRP (1:2,000) (C). Sera from a pre- and post-immunized rabbit were diluted 1:10,000 and assessed for anti-RABV NP polyclonal antibody production by immunoblotting (D). All gels used for immunoblotting were loaded with 10 ng of RABV NP per well. Molecular mass markers were Mark 12 or See Blue Plus 2 (Invitrogen).
Fig 3.
Comparison of RIADacetone test with FAT (inset) on canine brain smears infected with rabies virus (positive) or not (negative).
The presence of RABV antigen is indicated by brick red deposits (Positive) or green fluorescence (inset) in brain smears tested using the RIADacetone test or FAT, respectively. Magnification of the RIADacetone images are 63X and the FAT image is 20X. All smears were fixed in acetone.
Table 1.
Posterior medians and 95% probability intervals (PI) for diagnostic sensitivity (DSe) and specificity (DSp) from a 3-tests-in-2-population Bayesian latent class model.
Table 2.
Proficiency testing (PT) data using the RIADacetone test on 6 samples (4 positive and 2 negative for rabies infection) with 16 participating laboratories (A—P).
Table 3.
Combinations of test results by RIADformalin, RIADacetone, FAT and SST from brain smears from 116 suspected rabies infected dogs in Indonesia.