Neuraminidase inhibitor susceptibility testing in human influenza viruses: a laboratory surveillance perspective

Abstract

Neuraminidase inhibitors (NAIs) are vital in managing seasonal and pandemic influenza infections. NAI susceptibilities of virus isolates (n = 5540) collected during the 2008-2009 influenza season were assessed in the chemiluminescent neuraminidase inhibition (NI) assay. Box-and-whisker plot analyses of log-transformed IC(50)s were performed for each virus type/subtype and NAI to identify outliers which were characterized based on a statistical cutoff of IC(50) >3 interquartile ranges (IQR) from the 75(th) percentile. Among 1533 seasonal H1N1 viruses tested, 1431 (93.3%) were outliers for oseltamivir; they all harbored the H275Y mutation in the neuraminidase (NA) and were reported as oseltamivir-resistant. Only 15 (0.7%) of pandemic 2009 H1N1 viruses tested (n = 2259) were resistant to oseltamivir. All influenza A(H3N2) (n = 834) and B (n = 914) viruses were sensitive to oseltamivir, except for one A(H3N2) and one B virus, with D151V and D197E (D198E in N2 numbering) mutations in the NA, respectively. All viruses tested were sensitive to zanamivir, except for six seasonal A(H1N1) and several A(H3N2) outliers (n = 22) which exhibited cell culture induced mutations at residue D151 of the NA. A subset of viruses (n = 1058) tested for peramivir were sensitive to the drug, with exception of H275Y variants that exhibited reduced susceptibility to this NAI. This study summarizes baseline susceptibility patterns of seasonal and pandemic influenza viruses, and seeks to contribute towards criteria for defining NAI resistance.

Keywords: Oseltamivir; pandemic H1N1; peramivir; seasonal influenza A and B; zanamivir.

Figure 1

Box-and-whisker plots showing distributions of log-transformed oseltamivir IC₅₀ values for seasonal influenza A(H1N1) (n = 1533), 2009 pandemic H1N1 (n = 2259), seasonal influenza A(H3N2) (n = 834), and seasonal influenza B (n = 914) virus isolates. The boxes represent the 25^th to 75^th percentiles, and horizontal lines within the box represent median values. The whiskers represent the lowest and highest value in the 25^th percentile minus 1.5IQR and 75^th percentile plus 1.5IQR regions, respectively.

Figure 2

Box-and-whisker plots showing distribution of log-transformed peramivir IC₅₀ values for seasonal influenza A(H1N1) (n = 235), 2009 pandemic H1N1 (n = 550), seasonal influenza A(H3N2) (n = 220), and seasonal influenza B (n = 52). The boxes represent the 25^th to 75^th percentiles, and horizontal lines within the box represent median values. The whiskers represent the lowest and highest value in the 25^th percentile minus 1.5IQR and 75^th percentile plus 1.5IQR regions, respectively.

Figure 3

Box-and-whisker plots showing distributions of log-transformed zanamivir IC₅₀ values for seasonal influenza A(H1N1) (n = 1533), 2009 pandemic H1N1 (n = 2259), seasonal influenza A(H3N2) (n = 834), and seasonal influenza B (n = 914) virus isolates. The boxes represent the 25^th to 75^th percentiles, and horizontal lines within the box represent median values. The whiskers represent the lowest and highest value in the 25^th percentile minus 1.5IQR and 75^th percentile plus 1.5IQR regions, respectively.

Figure 4

Box-and-whisker plots showing distributions of log-transformed IC₅₀ values for oseltamivir carboxylate and peramivir among oseltamivir-sensitive (wildtype H275) seasonal influenza A(H1N1) virus isolates (n = 102). The boxes represent the 25^th to 75^th percentiles, and horizontal lines within the box represent median values. The whiskers represent the lowest and highest value in the 25^th percentile minus 1.5IQR and 75^th percentile plus 1.5IQR regions, respectively.