Effectiveness of Influenza Vaccine for Preventing Influenza Hospitalization.

<p>a Among the 4 hospitals which used IRDTs that can detect A(H1N1)pdm09.</p><p>Effectiveness of Influenza Vaccine for Preventing Influenza Hospitalization.</p

Effectiveness of Influenza Vaccine for Children, by Vaccine Doses (6m/o-12y/o).

<p>a Among the 4 hospitals which used IRDTs that can detect H1N1pdm09.</p><p>Effectiveness of Influenza Vaccine for Children, by Vaccine Doses (6m/o-12y/o).</p

Effectiveness of Influenza Vaccine for Children in 2013/14 Influenza Season (N = 4727).

<p>a Adjusted for comorbidity (yes or no, except H1N1 analysis), area (north area, middle area, south of the Kanto region), months of onset.</p><p>b Adjusted for age (0–15 y/o).</p><p>c Adjusted for time tested after the onset (<12, 12–48 and >48 hours).</p><p>d Patients tested only >12 hours after onset.</p><p>Data for 3046 patients were available; 1499 for any influenza (A; 537, H1N1; 43, B; 962) and 1547 for influenza negative.</p><p>e Two hospitals have no information on comorbidity.</p><p>f Only four institutes used IRDTs that can detect A(H1N1)pdm09. One hospital had no information on comorbidity.</p><p>g Not analyzed because few patients developed influenza.</p><p>* (vaccinated/cases) [vaccinated/controls].</p><p>Effectiveness of Influenza Vaccine for Children in 2013/14 Influenza Season (N = 4727).</p

Effectiveness of Influenza Vaccine, by Phase.

<p>a Among four hospitals which used IRDTs that can detect A(H1N1)pdm09.</p><p>* (vaccinated/cases) [vaccinated/controls].</p><p>Effectiveness of Influenza Vaccine, by Phase.</p

Effectiveness of Influenza Vaccine in Children (6 m/o-12 y/o) According to Hospital (A to V).

<p>a Among four hospitals which used IRDTs that can detect A(H1N1)pdm09.</p><p>b Number of patients vaccinated.</p><p>c Number of patients unvaccinated.</p><p>d VE against any influenza and VE against influenza B were higher in the north area than in the middle or south area (Breslow-Day, p < 0.05).</p><p>Effectiveness of Influenza Vaccine in Children (6 m/o-12 y/o) According to Hospital (A to V).</p

Time after the onset of fever and the number of patients who were diagnosed by the RT-SmartAmp assay as positive for infection with the 2009 pdm influenza A(H1N1) virus.

<p>Time after the onset of fever and the number of patients who were diagnosed by the RT-SmartAmp assay as positive for infection with the 2009 pdm influenza A(H1N1) virus.</p

Detection of the 2009 pdm influenza A(H1N1) virus by RT-SmartAmp assay in the fatal case.

<p><b>A:</b> Nasopharyngeal swab samples were collected at 11, 28, and 52 hours after the onset of fever from the patient who was transferred by ambulance to the National Center for Global Health and Medicine. The 2009 pdm influenza A(H1N1) virus was immediately detected by the RT-SmartAmp assay as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030236#s4" target="_blank">Materials and Methods</a>. <b>B:</b> Chest radiography of the patient was taken at 11 and 28 hours after the onset of fever. <b>C:</b> Partial sequence of the HA segment of the 2009 pdm influenza A(H1N1) virus was analyzed after extraction of viral genome RNA from the swab samples. An arrow indicates the mutation that caused an amino acid substitution at 185 from aspartate to asparagine (N) in the HA protein.</p

Preparation of SmartAmp primers to detect the HA segment of the 2009 pdm influenza A(H1N1) virus.

<p>A. Mutation rate and difference score in the consensus sequence of the HA segment. Nucleotide sequences of the HA segment of 2009 pdm influenza A(H1N1) viruses were obtained from the NCBI Influenza Virus Resource database and aligned by using the MUSCLE program to gain the consensus sequence of the HA segment. The mutation rate at each base position was calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030236#s4" target="_blank">Materials and Methods</a>. The difference between 2009 pdm and seasonal A(H1N1) viruses was calculated at each position in the nucleotide sequence of the HA segments to gain the difference score. B: Comparison of data acquired in 2009 and 2011 as to the mutation rates in the HA segment of the 2009 pdm influenza A(H1N1) viruses.</p

Comparison of results detected by the rapid diagnosis kits and the RT-SmartAmp assay.

<p>As depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030236#pone.0030236.s003" target="_blank">Figure S3</a>, two swab samples were collected as one pair from each patient. Swab samples were separately used for the rapid influenza diagnostic tests and RT-SmartAmp assay. All the samples were transferred to RIKEN Yokohama Institute for sequence analysis and RT-Q-PCR detection.</p

RT-SmartAmp detection of the 2009 pdm influenza A(H1N1) virus with different dilutions as well as the cross activity with seasonal A(H1N1), seasonal A(H3N2), and B(Victoria) viruses.

<p>Isolated and cultured influenza viruses, <i>i.e.</i>, 2009 pdm A(H1N1), seasonal A(H1N1), seasonal A(H3/N2), and seasonal B/Victoria, were prepared at a viral titer of 10⁷ pfu/ml. Each viral sample (10 µl), except for the 2009 pdm influenza A(H1N1) virus, was mixed with 90 µl of the pretreatment medium (5% SDS) to dissolve the viral membrane and to facilitate viral RNA extraction. A sample (15 µl) of the resulting medium was subjected to spin column chromatography, and the eluted solution (5 µl) was applied to the RT-SmartAmp reaction mixture. In the case of the 2009 pdm influenza A(H1N1) virus, the viral sample was diluted by 10³-, 10⁴-, or 10⁵-fold as indicated in the figure, and then processed in the same manner as described above. The RT-SmartAmp assay was performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030236#s4" target="_blank">Materials and Methods</a>.</p