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采用滤垫设计开发一种用于检测猪粪便样本中猪流行性腹泻病毒(PEDV)的准确侧向流动免疫分析方法。

Development of an accurate lateral flow immunoassay for PEDV detection in swine fecal samples with a filter pad design.

作者信息

Zou Siyi, Wu Lei, Li Gan, Wang Juan, Cao Dongni, Xu Tao, Jia Aiqing, Tang Yong

机构信息

Department of Bioengineering, Guangdong Province Engineering Research Center of Antibody Drug and Immunoassay, Jinan University, Guangzhou, 510632 China.

Guangdong Haid Institute of Animal Husbandry & Veterinary, Guangzhou, 511400 China.

出版信息

Anim Dis. 2021;1(1):27. doi: 10.1186/s44149-021-00029-1. Epub 2021 Nov 8.

Abstract

UNLABELLED

Porcine epidemic diarrhea virus (PEDV), as the main causative pathogen of viral diarrhea in pigs, has been reported to result in high morbidity and mortality in neonatal piglets and cause significant economic losses to the swine industry. Rapid diagnosis methods are essential for preventing outbreaks and transmission of this disease. In this study, a paper-based lateral flow immunoassay for the rapid diagnosis of PEDV in swine fecal samples was developed using stable color-rich latex beads as the label. Under optimal conditions, the newly developed latex bead-based lateral flow immunoassay (LBs-LFIA) attained a limit of detection (LOD) as low as 10 TCID/mL and no cross-reactivity with other related swine viruses. To solve swine feces impurity interference, by adding a filtration unit design of LFIA without an additional pretreatment procedure, the LBs-LFIA gave good agreement (92.59%) with RT-PCR results in the analysis of clinical swine fecal samples ( = 108), which was more accurate than previously reported colloidal gold LFIA (74.07%) and fluorescent LFIA (86.67%). Moreover, LBs-LFIA showed sufficient accuracy (coefficient of variance [CV] < 15%) and stable (room temperature storage life > 56 days) performance for PEDV detection, which is promising for on-site analysis and user-driven testing in pig production system.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1186/s44149-021-00029-1.

摘要

未标记

猪流行性腹泻病毒(PEDV)作为猪病毒性腹泻的主要致病病原体,据报道可导致新生仔猪的高发病率和死亡率,并给养猪业造成重大经济损失。快速诊断方法对于预防该疾病的爆发和传播至关重要。在本研究中,使用稳定的富含颜色的乳胶珠作为标记物,开发了一种用于快速诊断猪粪便样本中PEDV的基于纸的侧向流动免疫分析方法。在最佳条件下,新开发的基于乳胶珠的侧向流动免疫分析方法(LBs-LFIA)的检测限(LOD)低至10 TCID/mL,且与其他相关猪病毒无交叉反应。为了解决猪粪便杂质干扰问题,通过在不进行额外预处理程序的情况下添加LFIA的过滤单元设计,LBs-LFIA在分析临床猪粪便样本(n = 108)时与RT-PCR结果具有良好的一致性(92.59%),比先前报道的胶体金LFIA(74.07%)和荧光LFIA(86.67%)更准确。此外,LBs-LFIA在PEDV检测中显示出足够的准确性(变异系数[CV]<15%)和稳定的性能(室温储存寿命>56天),这对于猪生产系统中的现场分析和用户驱动检测具有前景。

补充信息

在线版本包含可在10.1186/s44149-021-00029-1获取的补充材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1185/8572657/13596ae99181/44149_2021_29_Fig1_HTML.jpg

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