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A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

文献类型： 外文期刊

作者： Huo, Caiyun 1 ; Li, Donghai 1 ; Hu, Zhenguo 1 ; Li, Guiping 1 ; Hu, Yanxin 2 ; Sun, Huiling 1 ;

作者机构： 1.Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing Key Lab Prevent & Control Infect Dis Live, Beijing, Peoples R China

2.China Agr Univ, Coll Vet Med, Key Lab Anim Epidemiol, Minist Agr, Beijing, Peoples R China

关键词： Avibacterium paragallinarum; lateral flow assay; nucleic acid detection; rapid; sensitive

期刊名称：FRONTIERS IN VETERINARY SCIENCE （ 影响因子：3.412； 五年影响因子：3.588 ）

ISSN：

年卷期： 2021 年 8 卷

页码：

收录情况： SCI

摘要： Avibacterium paragallinarum, the pathogen of infectious coryza, caused a highly contagious respiratory disease that poses a serious threat to chickens. Hence, it is necessary to do diagnostic screening for Av. paragallinarum. Existing technologies have been used for Av. paragallinarum testing, which, however, have some drawbacks such as time consuming and expensive that require well-trained personnel and sophisticated infrastructure, especially when they are limitedly feasible in some places for lack of resources. Nucleic acid hybridization-based lateral flow assay (LFA) is capable of dealing with these drawbacks, which is attributed to the advantages, such low cost, rapid, and simple. However, nucleic acid determination of Av. paragallinarum through LFA method has not been reported so far. In this study, we developed a novel LFA method that employed gold nanoparticle probes to detect amplified Av. paragallinarum dsDNA. Compared with agarose gel electrophoresis, this LFA strip was inexpensive, simple- to- use, and time- saving, which displayed the visual results within 5-8 min. This LFA strip had higher sensitivity that achieved the detection limit of 10(1) CFU/ml compared with 10(2) CFU/ml in agarose gel electrophoresis. Besides, great sensitivity was also shown in the LFA strip, and no cross reaction existed for other bacteria. Furthermore, Av. paragallinarum in clinical chickens with infectious coryza were perfectly detected by our established LFA strip. Our study is the first to develop the LFA integrated with amplification and sample preparation techniques for better nucleic acid detection of Av. paragallinarum, which holds great potential for rapid, accurate, and on-site determination methods for early diagnosis of Av. paragallinarum to control further spreading.

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