Study on Nucleic Acid Detection Method of African Swine Fever Virus Based on PCR-LFIA

XIANG Si-yi; SU Xiao-na; ZHANG Yong-yi; CHEN Jun-jie; YANG Xu-qiong; SHEN Yu-dong; YANG Jin-yi

doi:10.12452/j.fxcsxb.24012901

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Study on Nucleic Acid Detection Method of African Swine Fever Virus Based on PCR-LFIA

Experimental Techniques and Methods | 更新时间：2024-06-11

- Study on Nucleic Acid Detection Method of African Swine Fever Virus Based on PCR-LFIA
- Journal of Instrumental Analysis Vol. 43, Issue 6, Pages: 928-932(2024)
- 作者机构：
  
  华南农业大学食品学院广东省食品质量安全重点实验室，广东广州 510642
- 作者简介：
- 基金信息：
- DOI：10.12452/j.fxcsxb.24012901
  CLC： O657.3;R446.6
- Published：15 June 2024，
  
  Received：29 January 2024，
  
  Revised：08 March 2024，
- 稿件说明：
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向四意,苏晓娜,张咏仪,陈俊杰,杨旭琼,沈玉栋,杨金易.基于PCR-LFIA的非洲猪瘟病毒核酸检测方法研究[J].分析测试学报,2024,43(06):928-932.

XIANG Si-yi,SU Xiao-na,ZHANG Yong-yi,CHEN Jun-jie,YANG Xu-qiong,SHEN Yu-dong,YANG Jin-yi.Study on Nucleic Acid Detection Method of African Swine Fever Virus Based on PCR-LFIA[J].Journal of Instrumental Analysis,2024,43(06):928-932.
向四意,苏晓娜,张咏仪,陈俊杰,杨旭琼,沈玉栋,杨金易.基于PCR-LFIA的非洲猪瘟病毒核酸检测方法研究[J].分析测试学报,2024,43(06):928-932. DOI： 10.12452/j.fxcsxb.24012901.

XIANG Si-yi,SU Xiao-na,ZHANG Yong-yi,CHEN Jun-jie,YANG Xu-qiong,SHEN Yu-dong,YANG Jin-yi.Study on Nucleic Acid Detection Method of African Swine Fever Virus Based on PCR-LFIA[J].Journal of Instrumental Analysis,2024,43(06):928-932. DOI： 10.12452/j.fxcsxb.24012901.

摘要

该文利用聚合酶链式反应（PCR）技术进行扩增后，以侧流免疫层析方法（LFIA）对核酸进行检测，建立了一种非洲猪瘟病毒核酸的快速检测方法。通过设计引物，构建了pUC57-ASFV-（1-1941）质粒作为阳性质控品，采用柠檬酸三钠还原法制备胶体金颗粒用于标记抗体建立免疫分析方法。通过优化实验条件，在质粒浓度1.6×10

-2

~1.6×10

copies/μL范围内，得到pUC57-ASFV-（1-1941）阳性质粒的检出限为1.6

copies/μL。该方法与其他猪病毒无交叉，特异性良好，可作为非洲猪瘟现场筛查的辅助方法。

Abstract

In this paper，polymerase chain reaction（PCR） was used to amplify nucleic acid，lateral flow immunoassay（LFIA） was used to detect nucleic acid，and a rapid detection method for African swine fever virus nucleic acid was established. In this study，by designing primers，the positive plasmid pUC57-ASFV-（1-1941） was constructed as a quality control product and colloidal gold particles were prepared by trisodium citrate reduction method for antibody labeling to establish an immunoassay method. Under the optimal experimental conditi

ons，the plasmid concentration was in the range of 1.6×10

-2

-1.6×10

copies/μL，the detection limit of pUC57-ASFV-（1-1941） positive plasmid was 1.6 copies/μL，the method had no cross with other pig viruses and had good specificity.The established method can be used as an auxiliary method for field screening of African swine fever.

关键词

非洲猪瘟聚合酶链式反应（PCR）免疫层析可视化

Keywords

African swine feverpolymerase chain reaction（PCR）immunochromatographyvisualization

references

Tignon M，Gallardo C，Iscaro C，Hutet E，Van der Stede Y，Kolbasov D，De Mia G M，Le Potier M，Bishop R P，Arias M，Koenen F. J. Viro. Methods，2011，178（1/2）：161-170.

Galindo I，Alonso C. Viruses，2017，9（5）：103.

Montgomery R E. J. Comp. Pathol. Ther.，1921，34：159-191.

Zhou X，Li N，Luo Y，Liu Y，Miao F，Chen T，Zhang S F，Cao P L，Li X D，Tian K G，Qiu H J，Hu R L. Transboundary Emerging Dis.，2018，65（6）：1482-1484.

Gallardo C，Soler A，Nurmoja I，Cano-Gomez C，Cvetkova S，Fran M，Wozniakowski G，Simon A，Perez C R N，Arias M. Transboundary Emerging Dis.，2021，68（5）：2826-2841.

Yang Z S，Jiang Y N，Cheng N，Li X Y，Huang K L，Liu Q L，Sun Y L，Xu W T. J. Instrum. Anal. 杨湛森，蒋雅楠，程楠，李相阳，黄昆仑，刘清亮，孙艳丽，许文涛．分析测试学报），2021，40（5）：628-638.

World Organization for Animal Health（OIE）. African Swine Fever（Infection with African Swine Fever Virus）. ［2024-03-06］. https：//www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.09.01_ASF.pdfhttps://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.09.01_ASF.pdf.

Maffert P，Reverchon S，Nasser W，Rozand C，Abaibou H. Eur. J. Clin. Microbiol. Infect. Dis.，2017，36：1717-1731.

Yang H，Peng Z， Song W B，Zhang C，Fan J，Chen H J，Hua L，Pei J，Tang X B，Chen H C，Wu B. Front. Veter. Sci.，2022：9943099.

He J X，Ding X Y，Zhou X Y，Zhou Y X，Quan W X，Zhao J，Lin Z H，Li J T. J. Pathog. Biol.（何久香，丁晓艳，周晓杨，周伃欣，全纹萱，赵镜，林治华，李晋涛.中国病原生物学杂志），2022，17（5）：497-501，508.

Wu K K，Liu J M，Wang L X，Fan S Q，Li Z Y，Li Y W，Yi L，Ding H X，Zhao M Q，Chen J D. Vaccines，2020，8（3）：531-531.

Xia X H. Development and Primary Application of PCR-ELISA Method for the Detection of Mycoplasma Suis. Yanbian：Yanbian University（夏晓辉. 猪附红细胞体PCR-ELISA检测方法的建立及初步应用. 延边：延边大学），2013.

Cai Y，Zhang S N，Zhang J X，Liu X N，Ma K，Xu W，Deng X H，Yang J C，Ma T，Jiang C，Hui W L，Cui Y L. Anal. Chem.，2022，94：4686-4694.

Huang X L，Aguilar Z P，Xu H Y，Lai W H，Xiong Y H. Biosens. Bioelectron.，2016，75：166-180.

Sastre P，Gallardo C，Monedero A，Ruiz T，Arias M，Sanz A，Rueda P. BMC Veter. Res.，2016，12（1）：206.

GB/T 18648-2020. Diagnostic Techniques for African Swine Fever. National Standards of the People's Republic of China（非洲猪瘟诊断技术. 中华人民共和国国家标准）.

Yu Y Y，Zhang S L，Zou Y H，Qiu Y L. J. Instrum. Anal.（余宇燕，张淑玲，邹艳辉，邱亚利. 分析测试学报），2014，33（7）：830-834.

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