Development of a focus reduction neutralization test (FRNT) for detection of mumps virus neutralizing antibodies

作者：Vaidya Sunil R^*; Brown David W G; Jin Li; Samuel Dhanraj; Andrews Nick; Brown Kevin E

来源：Journal of Virological Methods, 2010, 163(1): 153-156.

DOI：10.1016/j.jviromet.2009.09.006

• Mumps neutralizing antibody
• Focus reduction neutralization test (FRNT)
• Assay validation

Although the plaque reduction neutralization test (PRNT) is considered the "gold-standard" assay for measuring neutralizing antibodies for mumps, it is technically demanding, slow and requires large serum volumes, which limits its use for investigating mumps vaccine efficacy and population susceptibility. Therefore, an immunocolourimetric-based focus reduction neutralization test (FRNT) was developed and validated against PRNT using 30 blood donor plasma samples (116 positive, 5 equivocal, and 9 negative for mumps IgG by EIA). The samples were tested in triplicate by FRNT and PRNT in 10 and 4 separate assay runs, respectively, and 50% neutralizing antibody titres calculated using the Karber formula. There was good correlation between the two neutralization assays (R(2) = 0.88). Inter-assay variation for FRNT titres was 2-fold, compared to a 3-fold variation for PRNT titres. From the distribution of results, a positive cut-off for FRNT was defined as 1:4. In conclusion, FRNT has similar sensitivity to the PRNT and offers the advantage of speed (2 days vs. 7 days), reduced sample volume (40 mu L vs. 150 mu L), and the possibility of automation using 96-well plates. FRNT appears to be a good substitute for PRNT for characterising the immune response to mumps and for vaccine efficacy studies.

• 出版日期2010-1