禽流感病毒H5、H9亚型RT-RAA-LFD检测方法的建立

doi:10.11843/j.issn.0366-6964.2025.12.033

Abstract

Abstract: Avian influenza (AI) is a significant threat to the healthy development of the poultry industry in China. It spreads rapidly and widely, with the potential for cross-species transmission, posing a severe threat to the health of both animals and humans. In particular, zoonotic influenza virus infections are highly hazardous and can cause substantial economic losses and social impacts. Therefore, the rapid and sensitive detection of common avian influenza viruses is of great significance for controlling the spread of the disease. This study aims to develop an efficient and convenient detection method for avian influenza, addressing the actual needs of prevention and control. Based on the conserved sequences of the H5N1 HA and H9N2 HA genes of the avian influenza virus (AIV), primers and probes were designed. A detection method for AIV using reverse transcription recombinase polymerase amplification (RT-RAA) combined with lateral flow dipstick (LFD) was established. This method can complete the detection in just 15 minutes at 37℃. There was no cross-reactivity between the AIV H5 subtype and the AIV H9 subtype, as well as between AIV and other viruses such as Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious bursal disease virus (IBDV), indicating good specificity. The sensitivity of the RT-RAA-LFD method can reach 100 copies per reaction, which is approximately 100 times higher than that of conventional RT-PCR. Furthermore, after testing 120 clinical samples, a 99% concordance rate was achieved between RT-RAA and commercialised RT-qPCR as well as conventional RT-PCR. The results demonstrate that this method can rapidly, accurately, conveniently, and intuitively detect AIV H5 and H9 subtypes, providing an effective and intuitive approach to the detection of avian influenza.

Key words: avian influenza virus, H5 subtype, H9 subtype, recombinase-aid amplification

CLC Number:

S852.659.5

LIN Yating, LIU Shuyi, QIU Deyang, YANG Dawei, CHEN Hu, LI Guimei, SHAO Di. Establishment of RT-RAA-LFD Detection Method for Avian Influenza Virus H5 and H9 Subtypes[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6308-6315.

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Establishment of RT-RAA-LFD Detection Method for Avian Influenza Virus H5 and H9 Subtypes

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