Volume 30, Number 8—August 2024
Dispatch
Multiplex Dual-Target Reverse Transcription PCR for Subtyping Avian Influenza A(H5) Virus
Malaya K. Sahoo, Ingrid E.A. Morante, ChunHong Huang, Daniel Solis, Fumiko Yamamoto, Uzoamaka C. Ohiri, Daniel Romero, and Benjamin A. Pinsky
Table 2
Avian influenza A(H5) virus genomic RNA used to evaluate inclusivity in study of multiplex dual-target reverse transcription PCR for subtyping avian influenza A(H5) virus*
| Genome | Clade | GenBank accession no. | H5 Ct | M Ct | Ct difference |
|---|---|---|---|---|---|
| Kilbourne F181, A/duck/Singapore/645/1997 (H5N3) | EA-non-Gs/Gd | NA | 18.3 | 17.7 | 0.6 |
| A/quail/California/14-012546-1/2014 (H5N8) | Am-non-Gs/Gd | NA | 22.9 | 19.0 | 3.9 |
| A/mallard/Minnesota/16-041335-3/2016 (H5N2) | Am-non-Gs/Gd | MH546659 | 32.1 | 22.3 | 9.9 |
| A/emperor goose/Alaska/17-004479-1/2016 (H5N2) | Am-non-Gs/Gd | MH546451 | 26.8 | 23.3 | 3.6 |
| A/glaucous-winged gull/Alaska/16-041335-19/2016 (H5N2) | Am-non-Gs/Gd | MH546475 | 23.3 | 21.8 | 1.5 |
| A/northern pintail/Alaska/16-041335-5/2016 (H5N2) | Am-non-Gs/Gd | MH546883 | 20.9 | 19.1 | 1.9 |
*Kilbourne F181 genomic RNA was obtained from BEI Resources (https://www.beiresources.org). Am-non-Gs/GD A(H5) genomic RNA was obtained from the US Department of Agriculture. The genomic RNA from these influenza A(H5) viruses was diluted 1:10 buffer AVE plus carrier RNA (QIAGEN, https://www.qiagen.com) and tested in duplicate. Ct values are means. Am-non-Gs/Gd, non-goose/GuangDong from the United States; Ct, cycle threshold; EA-non-Gs/Gd, Eurasian non-goose/GuangDong; M, matrix gene; NA, not available.
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