Comparative evaluation of some techniques used for the detection of rabies virus

Abstract

Background: Neurotropic viruses in the family Rhabdoviridae, genus Lyssavirus, are what cause rabies, an acute,
progressive, and highly lethal encephalomyelitis.
Aim: Evaluation of the used diagnostic techniques to determine the most simple; rapid and accurate test for rabies virus (RABV) recognition in different specimens aiming to reach a rapid diagnosis as a step aid in the disease control and to prevent or even minimize the suspected hazard.
Method: The used techniques included an infection trial of Swiss mice with the mice-adapted challenge rabies virus followed by the detection of the virus in the infected mices’ brains. Virus detection was carried out through the  application BHK21 cell line infection; fluorescent antibody technique; latex agglutination test (LAT); direct enzymelinked  immunosorbent assay (ELISA); rabies antigen detection kit ELISA; conventional polymerase chain reaction (PCR).   Results: It was found that virus inoculation in mice and BHK21 cell lines needs 5–7 days with positivity of 90% and
100%, respectively. Rapid antigen kit was able to detect rabies antigen in mice brains suspension and BHK21 infected
fluid within 3–5 minutes with percentages of 60% and 55.5%, respectively. In 1–1.5 hours, the direct fluorescent antibody method (DFAT) detected 90% and 100% of the rabies antigen in BHK21 cell line infection and brain impressions,  respectively. Latex agglutination showed clear results with 88.8% with BHK21 infected fluid within 3–5 minutes while it  did not carry out on brain emulsions to prevent falsely positive results brought on by the presence of tissue fragments.  Conventional one-step PCR revealed 100% positivity with either brain or cell culture preparations within 2 days. Direct  ELISA showed 88.8% positivity with BHK21 infected fluid with 1 day of work.
Conclusion: Mice inoculation test, cell culture infection; DFAT and PCR are the most accurate techniques for the detection of RABV with a positivity of 90%–100% followed by LAT and ELISA with a positivity of 88.8%, and lastly, rabies  antigen ELISA kit (RAK) with a positivity of 55.5%–60% taking in consideration the required time for each. In addition, the  positivity % of the applied tests revealed their sensitivity and specificity.