METHOD VALIDATION: QUANTITATION OF ANTI-HANTA VIRUS NEUTRALIZING ANTIBODIES BY THE VIRAL PLAQUE REDUCTION NEUTRALIZATION TEST

Abstract

The Hantavirus Plaque Reduction Neutralization Test (PRNT) used at the US Army Medical Research Institute of Infectious Diseases (USAMRIID) was validated by quantifying anti-Hantaan (HTNV) neutralizing antibodies (NAb) using applicable performance parameters and FY07-30 clinical sera. The assay demonstrated precision, accuracy, and linearity within the tested range by passing the acceptance criteria with ease. The limit of quantitation (LOQ) was revealed to be a PRNT50 titer of 20 and the agreed upon range was determined to start at the LOQ and go to an indefinite value. No statistical differences were observed when test samples were mixed with different matrices, indicative of a highly specific assay not easily affected by the presence of other components. Additionally, neither the cell passage number nor the source of FBS had a valid measurable effect on the assays robustness. Therefore this validation concludes that the hantavirus PRNT is a reliable method for quantifying anti-HTNV NAbs.