Microfluidic-LAMP chip for the point-of-care detection of gene-deleted and wild-type African swine fever viruses and other four swine pathogens

Chihai Ji; Ling Zhou; Yonghui Chen; Xueen Fang; Yanhong Liu; Mengkan Du; Xiandong Lu; Qianniu Li; Heng Wang; Yuan Sun; Tian Lan; Jingyun Ma

doi:10.3389/fvets.2023.1116352

Microfluidic-LAMP chip for the point-of-care detection of gene-deleted and wild-type African swine fever viruses and other four swine pathogens

Front Vet Sci. 2023 Feb 16:10:1116352. doi: 10.3389/fvets.2023.1116352. eCollection 2023.

Authors

Chihai Ji^{1

2

3}, Ling Zhou^{1

2}, Yonghui Chen^{1

2}, Xueen Fang⁴, Yanhong Liu⁵, Mengkan Du⁶, Xiandong Lu⁵, Qianniu Li^{1

2}, Heng Wang^{2

3}, Yuan Sun^{1

2}, Tian Lan^{1

2}, Jingyun Ma^{1

2}

Affiliations

¹ Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, Guangdong, China.
² Guangdong Laboratory for Lingnan Modern Agriculture, College of Animal Science, South China Agricultural University, Guangzhou, China.
³ African Swine Fever Regional Laboratory of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
⁴ Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
⁵ Ningbo iGene Technology Co., Ltd., Ningbo, China.
⁶ Hangzhou Xiaoshan District Animal Husbandry and Veterinary Development Center, Xiaoshan Bureau of Animal Husbandry and Veterinary, Hangzhou, China.

Abstract

Introduction: Different pathogens causing mixed infection are now threatening the pig industry in the context of the African Swine Fever (ASF) circulating especially in China, and it is crucial to achieving the early diagnosis of these pathogens for disease control and prevention.

Methods: Here we report the development of a rapid, portable, sensitive, high-throughput, and accurate microfluidic-LAMP chip detection system for simultaneous detection and differentiation of gene-deleted type and wild-type African swine fever virus (ASFV), pseudorabie virus (PRV), porcine parvovirus (PPV), porcine circovirus type 2 (PCV2), and porcine reproductive and respiratory syndrome (PRRSV).

Results and discussion: The newly developed system was shown to be sensitive with detection limits of 101 copies/μl for ASFV-MGF505-2R/P72, PPV, and PCV2, 102 copies/μl for ASFV-CD2v, PRV, and PRRSV. The system was highly specific (100%) and stable (C.V.s < 5%) in its ability to detect different pathogens. A total 213 clinical samples and 15 ASFV nucleic acid samples were collected to assess the performance of the detection system, showing highly effective diagnosis. Altogether, the developed microfluidic-LAMP chip system provides a rapid, sensitive, high-throughput and portable diagnostic tool for the accurate detection of multiple swine pathogens.

Keywords: loop-mediated isothermal amplification (LAMP); microfluidic chip; multiplex detection; pathogens; portable diagnostic tool.

Grants and funding

In support of this work, the Provincial Key Field Research and Development Plan funded the Research and Application of Accurate Detection Technology of African Swine Fever (No. 201902112019B020211005), the Guangdong Major Project of Basic and Applied Basic Research (No. 2020B0301030007), and the Natural Science Foundation of Guangdong Province (Nos. 2022A1515012473, 2020A1515010295, and 2020A1515010220).