适合早期诊断的非洲猪瘟sIgA抗体量子点免疫层析检测方法建立

doi:10.11843/j.issn.0366-6964.2023.10.027

Abstract

Abstract: African swine fever (ASF), caused by the African swine fever virus (ASFV), is a devastating infectious disease in wild boars and domestic pigs. The extremely high infectivity and morbidity of ASFV have caused enormous socioeconomic losses. Given that there is no effective vaccine available for ASFV infection, early diagnosis is crucial for the prevention and control of ASF. Therefore, it is of great significance to develop a sensitive and rapid method for the on-site detection of ASFV. Firstly, the detection probe of quantum dot microspheres (QDM) coupled ASFV recombinant antigen p30 and the quality control probe of QDM coupled anti-chicken IgY were prepared based on the condensation reaction between amino and carboxyl groups. Then, the immunostrip was prepared by immobilized anti-porcine IgA-Sc antibody and chicken IgY protein on the detection line and control line, respectively. The ASFV-specific sIgA antibody in the oral fluid can be captured by the QDM-p30 fixed on the T-line. Finally, the detection sensitivity was tested by detecting the ASFV-positive oral fluid with double gradient dilution; the specificity was verified by detecting several other common swine disease viruses; and the clinical diagnostic efficacy was evaluated by testing clinical oral fluid samples from ASFV-negative and ASFV-positive animals. The proposed method shows good specificity and no cross-reaction with PRV, CSFV and PRRV; shows high sensitivity with the lowest detected dilution of 1:64 for ASFV-positive oral fluid; shows short time consuming within 20 minutes; and is easy to operate without intrusive sampling. In addition, sIgA-positive conversion could be detected as early as the 6th day after ASFV infection, suggesting the potential for early diagnosis. Here, we developed a QDM-based immunostrip for on-site early detection of ASFV sIgA antibody in oral fluid, which provides new technical support and supplement for ASFV monitoring, prevention and control.

Key words: African swine fever, sIgA antibody, QDMs-based immunostrip, point-of care testing

CLC Number:

S858.285.3

XIE Qingyun, YI Weijie, LI Jiahao, BAI Yun, XIE Xing, YUAN Ting, ZHANG Yue, FENG Yufan, ZHAO Dongming, BU Zhigao, LIU Fei, FENG Zhixin. Development of Quantum Dot Microsphere-based Immunostrip for Early Detection of Specific sIgA Antibody to African Swine Fever Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4311-4319.

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Development of Quantum Dot Microsphere-based Immunostrip for Early Detection of Specific sIgA Antibody to African Swine Fever Virus

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