Abstract:
Acute hepatopancreatic necrosis disease (AHPND) poses a significant threat to shrimp aquaculture worldwide, necessitating the accurate and rapid detection of the pathogens. However, the increasing number of Vibrio species that cause the disease makes diagnosis and control more difficult. This study focuses on developing a monoclonal antibody against the Photorhabdus insect-related (Pir) toxin B (PirB), a pivotal virulence factor in AHPND-causing Vibrio, and establishing a colloidal gold immunochromatographic assay for the enhanced early diagnosis and monitoring of AHPND. Monoclonal antibodies targeting PirB were developed and utilized in the preparation of colloidal-gold-labeled antibodies for the immunochromatographic assay. The specificity and sensitivity of the assay were evaluated through various tests, including antibody subclass detection, affinity detection, and optimal labeling efficiency assessment. The developed PirB immunochromatographic test strips exhibited a good specificity, as demonstrated by the positive detection of AHPND-causing Vibrio and negative results for non-AHPND-causing Vibrio. The study highlights the potential of the developed monoclonal antibody and immunochromatographic assay for the effective detection of AHPND-causing Vibrio. Further optimization is needed to enhance the sensitivity of the test strips for improved practical applications in disease prevention and control in shrimp aquaculture.
摘要:
急性肝胰腺坏死病(AHPND)对世界范围内的对虾养殖构成重大威胁,需要准确和快速地检测病原体。然而,引起该疾病的弧菌种类的增加使得诊断和控制更加困难。这项研究的重点是开发针对光纹肌昆虫相关(Pir)毒素B(PirB)的单克隆抗体,引起AHPND的弧菌中的关键毒力因子,并建立胶体金免疫层析法,用于增强AHPND的早期诊断和监测。开发了靶向PirB的单克隆抗体,并将其用于制备胶体金标记的抗体,用于免疫层析测定。通过各种测试评估该测定的特异性和敏感性,包括抗体亚类检测,亲和力检测,和最佳标签效率评估。所研制的PirB免疫层析试纸条具有良好的特异性,如AHPND引起的弧菌的阳性检测和非AHPND引起的弧菌的阴性结果所证明的。该研究强调了开发的单克隆抗体和免疫层析测定法用于有效检测引起AHPND的弧菌的潜力。需要进一步优化以增强测试条的灵敏度,以改善对虾水产养殖中疾病预防和控制的实际应用。