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. 2023 Apr;95(4):e28700.
doi: 10.1002/jmv.28700.

Performance assessment and validation of a plaque reduction neutralization test (PRNT) in support to yellow fever diagnostic and vaccine clinical trials

Moussa Dia  1 , Ndeye Sakha Bob  1 , Cheikh Talla  2 , Anne Dupressoir  3 , Camille Escadafal  4 , Mareme Seye Thiam  2 , Amadou Diallo  2 , Oumar Ndiaye  1 , Jean Michel Heraud  1 , Oumar Faye  1 , Amadou Alpha Sall  1 , Ousmane Faye  1 , Gamou Fall  1
Affiliations

Performance assessment and validation of a plaque reduction neutralization test (PRNT) in support to yellow fever diagnostic and vaccine clinical trials

Moussa Dia et al. J Med Virol. 2023 Apr.

Abstract

Yellow fever (YF) virus is a mosquito-borne virus belonging to the Flaviviridae family that circulates in tropical and subtropical areas of Africa and South America. Despite the availability of an effective vaccine, YF remains a threat to travelers, residents of endemic areas, and unvaccinated populations. YF vaccination and natural infection both induce the production of neutralizing antibodies. Serological diagnostic methods detecting YF virus-specific antibodies demonstrate high levels of cross-reactivities with other flaviviruses. To date, the plaque reduction neutralization test (PRNT) is the most specific serological test for the differentiation of flavivirus infections and is considered the reference method for detecting YF neutralizing antibodies and assessing the protective immune response following vaccination. In this study, we developed and validated a YF PRNT. We optimized different parameters including cell concentration and virus-serum neutralization time period and then assessed the intra- and inter-assay precisions, dilutability, specificity, and lower limit of quantification (LLOQ) using international standard YF serum, sera from vaccinees and human specimens collected through YF surveillance. The YF PRNT has shown good robustness and 100% of intra-assay precision, 95.6% of inter-assay precision, 100% of specificity, 100% of LLOQ, and 95.3% of dilutability. The test is, therefore, suitable for use in the YF diagnostic as well as evaluation of the YF vaccine neutralizing antibody response and risk assessment studies.

Keywords: PRNT; diagnostic and vaccine efficacy; neutralizing antibodies; precision; sensitivity; specificity; yellow fever.

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