Portable lab-on-a-chip platform enabling multiplex recombinant enzyme polymerase amplification detection of H5/H7/H10 avian influenza virus subtypes
文献类型： 外文期刊
作者： Liang, Li-guo 1 ; Wang, Ping 1 ; Fu, Jiamin 1 ; Zhu, Linwei 1 ; Cheng, Linfang 1 ; Liu, Fumin 1 ; Wu, Nanping 1 ; Xu, Lihua 4 ; Yao, Hangping 1 ; Wu, Haibo 1 ;
作者机构： 1.Zhejiang Univ, State Key Lab Diag & Treatment Infect Dis, 79 Qingchun Rd, Hangzhou 310003, Zhejiang, Peoples R China
2.Zhejiang Univ, Affiliated Hosp 1, Natl Clin Res Ctr Infect Dis, Sch Med, 79 Qingchun Rd, Hangzhou 310003, Zhejiang, Peoples R China
3.Zhejiang Chinese Med Univ, Affiliated Hosp 1, Ctr Clin Lab, Hangzhou 310006, Peoples R China
4.Zhejiang Acad Agr Sci, Anim Husb & Vet Inst, Hangzhou 310021, Peoples R China
关键词： Avian influenza virus; Subtype; Detection; Microfluidic microarray; Recombinant enzyme polymerase amplification
期刊名称：POULTRY SCIENCE （ 影响因子：4.2； 五年影响因子：4.5 ）
ISSN： 0032-5791
年卷期： 2025 年 104 卷 9 期
页码：
收录情况： SCI
摘要： The zoonotic nature of influenza pathogens creates substantial health security risks, jeopardizing the welfare of interconnected human and animal ecosystems. The H5/H7/H10 avian influenza virus (AIV) variants demonstrate persistent endemicity in poultry reservoirs and recurrent zoonotic jumps precipitating fatal human infections. Therefore, the innovation of multiplex diagnostic platforms integrating expedited processing, enhanced sensitivity, and subtype-specific discrimination has emerged as a pivotal strategy to curb epidemiological escalation. This research introduces a temperature-controlled nucleic acid detection platform utilizing micro-fluidic technology, enabling concurrent differentiation of H5, H7, and H10 AIV subtypes. Based on the conserved sequences of the hemagglutinin (HA) gene of H5, H7, and H10 AIVs, three sets of primers and probes specific to the subtypes were developed. These were then combined with microfluidic microarray technology and recombinant enzyme polymerase amplification. This combination aimed to create a method for the simultaneous detection of H5, H7, and H10 AIVs for differential diagnostic purposes. The method was distinguished by its specificity, sensitivity, accuracy, and its ability to detect these viruses in clinical samples. The specificity of the method showed that it could detect all strains of H5, H7 and H10 AIVs at the same time, with no cross-reactivity with other subtype influenza viruses or other avian pathogens. The sensitivity results showed that the assay could still detect the three AIV target genes simultaneously at a concentration of 2 copies per reaction. The results of this method for 100 clinical samples were consistent with those produced by quantitative PCR. This integrated detection system for H5/H7/H10 AIV differentiation exhibits exceptional specificity, enhanced sensitivity, rapid turnaround, and streamlined operational procedures, representing a viable solution for prompt pathogen identification during outbreaks.
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