Development of a reverse transcription recombinase polymerase amplification combined with lateral‐flow dipstick assay for avian influenza H9N2 HA gene detection

2018 ◽  
Vol 66 (1) ◽  
pp. 546-551 ◽  
Author(s):  
Zeng Wang ◽  
Pan‐Pan Yang ◽  
Yu‐Han Zhang ◽  
Kai‐Yue Tian ◽  
Chuan‐Zhou Bian ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Reverse Transcription ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Gene Detection ◽  
Ha Gene ◽  
Lateral Flow Dipstick ◽  
Avian Influenza H9n2

scholarly journals Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of Avian Influenza Virus H9N2 HA Gene

2021 ◽  
Vol 8 (7) ◽  
pp. 134
Author(s):  
Nahed Yehia ◽  
Fatma Eldemery ◽  
Abdel-Satar Arafa ◽  
Ahmed Abd El Wahed ◽  
Ahmed El Sanousi ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Reverse Transcription ◽  
Rapid Detection ◽  
Influenza A ◽  
Recombinase Polymerase Amplification ◽  
Economic Losses ◽  
Clinical Samples ◽  
Analytical Sensitivity ◽  
Rt Pcr ◽  
Ha Gene

The H9N2 subtype of avian influenza A virus (aIAV) is circulating among birds worldwide, leading to severe economic losses. H9N2 cocirculation with other highly pathogenic aIAVs has the potential to contribute to the rise of new strains with pandemic potential. Therefore, rapid detection of H9 aIAVs infection is crucial to control virus spread. A qualitative reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of aIAV subtype H9N2 was developed. All results were compared to the gold standard (real-time reverse transcription polymerase chain reaction (RT-PCR)). The RT-RPA assay was designed to detect the hemagglutinin (HA) gene of H9N2 by testing three pairs of primers and a probe. A serial concentration between 106 and 100 EID50 (50% embryo infective dose)/mL was applied to calculate the analytical sensitivity. The H9 RT-RPA assay was highly sensitive as the lowest concentration point of a standard range at one EID50/mL was detected after 5 to 8 min. The H9N2 RT-RPA assay was highly specific as nucleic acid extracted from H9 negative samples and from other avian pathogens were not cross detected. The diagnostic sensitivity when testing clinical samples was 100% for RT-RPA and RT-PCR. In conclusion, H9N2 RT-RPA is a rapid sensitive and specific assay that easily operable in a portable device for field diagnosis of aIAV H9N2.

scholarly journals Development of a reverse transcription recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of classical swine fever virus

2020 ◽  
Author(s):  
Jindai Fan ◽  
Yuanyuan Zhang ◽  
Wenxian Chen ◽  
Jingyuan Zhang ◽  
Chenchen Liu ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Professional Staff ◽  
Lateral Flow ◽  
Fever Virus ◽  
Recombinase Polymerase Amplification ◽  
Economic Losses ◽  
Simple Method ◽  
Lateral Flow Dipstick

Abstract Background: Classical swine fever (CSF), caused by the infection of Classical swine fever virus (CSFV), is a highly contagious disease of pigs and has caused significant economic losses in the pig industry. The rapid and effective detection of CSFV would contribute to the eradication program against CSF. Thus, this study aimed to develop a rapid and simple method for CSFV detection.Results: Here, a new method based on reverse transcription recombinase polymerase amplification (RT-RPA) coupled with lateral flow dipstick (LFD) was established for detecting CSFV. The RPA assay could be completed within 20 min at 37oC and the results of the RPA assay could be visualized by LFD assay with the naked-eye inspection. This RT-RPA-LFD assay could be used to detect CSFV specifically, with no cross-reaction with other pathogens. Its detection limit was 10 pg of CSFV cDNA. Importantly, the RT-RPA-LFD assay has a good performance on the CSFV detection for clinical samples.Conclusions: The established RT-RPA-LFD assay greatly reduced the need for professional staff and sophisticated instruments and made disease detection convenient and feasible. This method would be useful for the prevention and control of CSF, especially in resource-limited settings.

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