Abstract
BackgroundRabies is a major public health problem with a fatality rate close to 100%; however, complete prevention can be achieved through pre- or post-exposure prophylaxis. The rapid fluorescent focus inhibition test (RFFIT) is one of the recommended testing methods to determine the production of neutralizing antibodies after vaccination. Here, we report the development of a new monoclonal antibody (mAb) designed to react specifically with Rabies virus (RABV) phosphoprotein (P protein), and the evaluation of its applicability to the RFFIT and its effectiveness as a diagnostic reagent for human rabies.Methodology/principal findingsThe mAb KGH P 16B8 was produced to target the P protein of the Korean KGH RABV strain. An indirect immunofluorescence assay (IFA) was conducted to detect various strains of RABV in various cell lines. Alexa-conjugated KGH P 16B8 (16B8-Alexa) was developed for the RFFIT. The IFA test could detect RABV up to a 1:2,500 dilution, with a detection limit comparable to that of a commercial diagnostic reagent. The sensitivity, specificity, positive predictive value, and negative predictive value of the RFFIT using 16B8-Alexa in 414 clinical specimens were 98.67%, 99.47%, 99.55%, and 98.42%, respectively. The results of the RFFIT with 16B8-Alexa were strongly correlated with those obtained using an existing commercial diagnostic reagent (r = 0.995, p<0.001).Conclusions/significanceThe mAb developed in this study shows high sensitivity and specificity, confirming its clinical utility with the RFFIT to measure the rabies neutralizing antibody titer and establish a diagnosis in human. Thus, 16B8-Alexa is expected to serve as an alternative diagnostic reagent that is widely accessible, with potentially broad applications beyond those of the RFFIT in Korea. Further studies with 16B8-Alexa should provide insight into the immunological mechanism of the P protein of Korean RABV.
Highlights
Rabies is the oldest and most fatal viral zoonosis that has been known to humans for at least 2,300 years
Rabies represents a significant cause of fatality upon presentation of symptoms; pre- or timely post-exposure prophylaxis can provide complete protection to a population
The current reagents available for laboratory tests to determine the level of Rabies virus (RABV) neutralizing antibodies are not readily accessible in several regions, including Korea, and are associated with time and economic constraints owing to the import process
Summary
Rabies is the oldest and most fatal viral zoonosis that has been known to humans for at least 2,300 years. Rabies cases reappeared in 1993 in animals, followed by continuous case reports since . The complete genome sequence of the Rabies virus (RABV) KGH strain has been determined. KGH was the first human RABV strain isolated in Korea from hair follicles of a rabies patient whose symptoms developed following a raccoon bite in 2001. We report the development of a new monoclonal antibody (mAb) designed to react with Rabies virus (RABV) phosphoprotein (P protein), and the evaluation of its applicability to the RFFIT and its effectiveness as a diagnostic reagent for human rabies
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