Viral RNA Extraction and Quantification

Viral RNA from oropharyngeal and cloacal swabs was extracted from 200 μL of the supernatant using the MagNA Pure 96 extraction system (Roche, Manheim, Germany) according to the manufacturer’s instructions. The extracted RNA was quantified by real-time reverse transcription-polymerase chain reaction using previously described protocols (28 (link)). The Ct values were converted into infectious units equivalent to EID₅₀/ml using a standard curve.

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Lee S.H., Lee J., Noh J.Y., Jeong J.H., Kim J.B., Kwon J.H., Youk S., Song C.S, & Lee D.H. (2023). Age is a determinant factor in the susceptibility of domestic ducks to H5 clade 2.3.2.1c and 2.3.4.4e high pathogenicity avian influenza viruses. Frontiers in Veterinary Science, 10, 1207289.

Publication 2023

MagNA Pure 96 extraction system

Cloacal Infectious Oropharyngeal Real time polymerase chain reaction Reverse transcription Viral rna

Corresponding Organization : Konkuk University

Other organizations : Kyungpook National University, Chungbuk National University

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Variable analysis

independent variables

Viral RNA from oropharyngeal and cloacal swabs

dependent variables

Infectious units equivalent to EID50/ml

control variables

RNA extraction using the MagNA Pure 96 extraction system (Roche, Manheim, Germany) according to the manufacturer's instructions
Quantification of extracted RNA by real-time reverse transcription-polymerase chain reaction using previously described protocols (28)

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