tetano
Editor, Senior Moderator
Virus Genes
. 2023 Feb 13.
doi: 10.1007/s11262-023-01979-z. Online ahead of print.
Reverse transcription recombinase-aided amplification assay for avian influenza virus
Suchun Wang[SUP] #[/SUP][SUP] 1 2 [/SUP], Qingye Zhuang[SUP] #[/SUP][SUP] 1 3 [/SUP], Nan Jiang[SUP] 1 4 [/SUP], Fuyou Zhang[SUP] 1 [/SUP], Qiong Chen[SUP] 5 [/SUP], Ran Zhao[SUP] 5 [/SUP], Yang Li[SUP] 1 [/SUP], Xiaohui Yu[SUP] 1 [/SUP], Jinping Li[SUP] 1 [/SUP], Guangyu Hou[SUP] 1 [/SUP], Liping Yuan[SUP] 1 [/SUP], Fuliang Sun[SUP] 4 [/SUP], Zihao Pan[SUP] 6 [/SUP], Kaicheng Wang[SUP] 7 8 [/SUP]
Affiliations
Abstract
Avian influenza virus (AIV) infection can lead to severe economic losses in the poultry industry and causes a serious risk for humans. A rapid and simple test for suspected viral infection cases is crucial. In this study, a reverse transcription recombinase-aided amplification assay (RT-RAA) for the rapid detection of all AIV subtypes was developed. The reaction temperature of the assays is at 39 °C and the detection process can be completed in less than 20 min. The specificity results of the assay showed that this method had no cross-reaction with other main respiratory viruses that affect birds, including Newcastle disease virus (NDV) and infectious bronchitis virus (IBV). The analytical sensitivity at a 95% confidence interval was 10[SUP]2[/SUP] RNA copies per reaction. In comparison with a published assay for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), the κ value of the RT-RAA assay in 384 avian clinical samples was 0.942 (p < 0.001). The sensitivity and specificity of the RT-RAA assay for avian clinical sample detection was determined as 97.59% (95% CI 93.55-99.23%) and 96.79% (95% CI 93.22-98.59%), respectively. The RT-RAA assay for AIV in this study provided an effective and practicable tool for AIV molecular detection.
Keywords: Avian influenza virus; Reverse transcription recombinase-aided amplification assay; Sensitivity; Specificity.
. 2023 Feb 13.
doi: 10.1007/s11262-023-01979-z. Online ahead of print.
Reverse transcription recombinase-aided amplification assay for avian influenza virus
Suchun Wang[SUP] #[/SUP][SUP] 1 2 [/SUP], Qingye Zhuang[SUP] #[/SUP][SUP] 1 3 [/SUP], Nan Jiang[SUP] 1 4 [/SUP], Fuyou Zhang[SUP] 1 [/SUP], Qiong Chen[SUP] 5 [/SUP], Ran Zhao[SUP] 5 [/SUP], Yang Li[SUP] 1 [/SUP], Xiaohui Yu[SUP] 1 [/SUP], Jinping Li[SUP] 1 [/SUP], Guangyu Hou[SUP] 1 [/SUP], Liping Yuan[SUP] 1 [/SUP], Fuliang Sun[SUP] 4 [/SUP], Zihao Pan[SUP] 6 [/SUP], Kaicheng Wang[SUP] 7 8 [/SUP]
Affiliations
- PMID: 36781819
- DOI: 10.1007/s11262-023-01979-z
Abstract
Avian influenza virus (AIV) infection can lead to severe economic losses in the poultry industry and causes a serious risk for humans. A rapid and simple test for suspected viral infection cases is crucial. In this study, a reverse transcription recombinase-aided amplification assay (RT-RAA) for the rapid detection of all AIV subtypes was developed. The reaction temperature of the assays is at 39 °C and the detection process can be completed in less than 20 min. The specificity results of the assay showed that this method had no cross-reaction with other main respiratory viruses that affect birds, including Newcastle disease virus (NDV) and infectious bronchitis virus (IBV). The analytical sensitivity at a 95% confidence interval was 10[SUP]2[/SUP] RNA copies per reaction. In comparison with a published assay for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), the κ value of the RT-RAA assay in 384 avian clinical samples was 0.942 (p < 0.001). The sensitivity and specificity of the RT-RAA assay for avian clinical sample detection was determined as 97.59% (95% CI 93.55-99.23%) and 96.79% (95% CI 93.22-98.59%), respectively. The RT-RAA assay for AIV in this study provided an effective and practicable tool for AIV molecular detection.
Keywords: Avian influenza virus; Reverse transcription recombinase-aided amplification assay; Sensitivity; Specificity.