21029748
not annotated - annotated - LINNAEUS only
One-step real-time reverse transcription-PCR assays for detecting and subtyping pandemic influenza A/H1N1 2009, seasonal influenza A/H1N1, and seasonal influenza A/H3N2 viruses.
Pandemic influenza A/H1N1 2009 (A/H1N1pdm) virus has caused significant outbreaks worldwide. A previous one-step real-time reverse transcription-PCR (rRT-PCR) assay for detecting A/H1N1pdm virus (H1pdm rRT-PCR assay) was improved since the former probe had a low melting temperature and low tolerance to viral mutation. To help with the screening of the A/H1N1pdm virus, rRT-PCR assays were also developed for detecting human seasonal A/H1N1 (H1 rRT-PCR assay) and A/H3N2 influenza viruses (H3 rRT-PCR assay). H1pdm, H1, and H3 rRT-PCR assays were evaluated using in vitro-transcribed control RNA, isolated viruses, and other respiratory pathogenic viruses, and were shown to have high sensitivity, good linearity (R(2)=0.99), and high specificity. In addition, the improved H1pdm rRT-PCR assay could detect two viral strains of A/H1N1pdm, namely, A/Aichi/472/2009 (H1N1)pdm and A/Sakai/89/2009 (H1N1)pdm, which have mutation(s) in the probe-binding region of the hemagglutinin gene, without loss of sensitivity. Using the three rRT-PCR assays developed, 90 clinical specimens collected between May and October 2009 were then tested. Of these, 26, 20, and 2 samples were identified as positive for A/H1pdm, A/H3, and A/H1, respectively, while 42 samples were negative for influenza A viruses. The present results suggest that these highly sensitive and specific H1pdm, H1, and H3 rRT-PCR assays are useful not only for diagnosing influenza viruses, but also for the surveillance of influenza viruses.
Ann file
T1 Species 89 105 influenza A/H1N1
N1 Reference T1 Taxonomy:11320
T2 Species 121 137 influenza A/H1N1
N2 Reference T2 Taxonomy:11320
T3 Species 152 176 influenza A/H3N2 viruses
N3 Reference T3 Taxonomy:11320
T4 Species 189 205 influenza A/H1N1
N4 Reference T4 Taxonomy:11320
T5 Species 359 374 A/H1N1pdm virus
N5 Reference T5 Taxonomy:11320
T6 Species 534 549 A/H1N1pdm virus
N6 Reference T6 Taxonomy:11320
T7 Species 600 605 human
N7 Reference T7 Taxonomy:9606
T8 Species 615 621 A/H1N1
N8 Reference T8 Taxonomy:11320
T9 Species 645 669 A/H3N2 influenza viruses
N9 Reference T9 Taxonomy:11320
T10 Species 992 1018 viral strains of A/H1N1pdm
N10 Reference T10 Taxonomy:11320
T11 Species 1028 1054 A/Aichi/472/2009 (H1N1)pdm
N11 Reference T11 Taxonomy:11320
T12 Species 1059 1084 A/Sakai/89/2009 (H1N1)pdm
N12 Reference T12 Taxonomy:11320
T13 Species 1378 1385 A/H1pdm
N13 Reference T13 Taxonomy:11320
T14 Species 1387 1391 A/H3
N14 Reference T14 Taxonomy:11320
T15 Species 1397 1401 A/H1
N15 Reference T15 Taxonomy:11320
T16 Species 1452 1471 influenza A viruses
N16 Reference T16 Taxonomy:11320
T17 Species 1610 1627 influenza viruses
N17 Reference T17 Taxonomy:11309
T18 Species 1662 1679 influenza viruses
N18 Reference T18 Taxonomy:11309
#1 AnnotatorNotes T3 TODO check span: virus related
T19 Species 212 221 A/H1N1pdm
N19 Reference T19 Taxonomy:11320