Duong Ngoc-Diem, Nguyen-Phuoc Khai-Hoan, Mai-Hoang Thuy-Dung, Do Kim-Yen Thi, Huynh Tuan-Binh, Nguyen Nguyet-Thu Thi, Tran Thuoc Linh, Tran-Van Hieu
Laboratory of Biosensors, Faculty of Biology and Biotechnology, University of Science, 227 Nguyen Van Cu Street, Ward 4, District 5, Ho Chi Minh City, Vietnam.
Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh City, Vietnam.
3 Biotech. 2022 Oct;12(10):243. doi: 10.1007/s13205-022-03311-2. Epub 2022 Aug 24.
Acute hepatopancreatic necrosis disease (AHPND) is a contagious disease for the shrimp cultivation, thus early detection of disease is an unmet need. This present study documented for the first time a simple lateral flow immunoassay (LFIA) strip using polyclonal antibodies was created for the rapid detection both of PirA and PirB protein simultaneously. LFIA method based on the principle of sandwich format. The label is the colloidal gold. The polyclonal antibody was conjugated with the colloidal gold acting as biorecognition element and coated onto the conjugate pad. The rabbit anti-Pir, anti-PirB antibodies, and goat anti-rabbit IgG antibody were separately sprayed onto a nitrocellulose membrane to form two test lines and one control line, respectively. The appearance of red bands at the control line and the test line indicated a positive result. A single coloured band at control area indicated a negative result. The limit of detection of LFIA was found to be 125 ng, which could be visually detected by naked eye within 15 min. There was no cross-reactivity observed with VP. Furthermore, the sensitivity and specificity of LFIA were 94.0% and 98.0%, respectively. The developed test strip could be a game changer for early and in situ diagnosis of AHPND.
急性肝胰腺坏死病（AHPND）是一种对虾养殖具有传染性的疾病，因此疾病的早期检测是一个尚未满足的需求。本研究首次记录了一种使用多克隆抗体的简单侧向流动免疫分析（LFIA）试纸条，用于同时快速检测PirA和PirB蛋白。LFIA方法基于夹心形式的原理。标记物是胶体金。将多克隆抗体与作为生物识别元件的胶体金偶联，并包被在偶联垫上。将兔抗Pir、抗PirB抗体和山羊抗兔IgG抗体分别喷在硝酸纤维素膜上，分别形成两条检测线和一条对照线。对照线和检测线出现红色条带表示阳性结果。对照区域出现单一色带表示阴性结果。发现LFIA的检测限为125 ng，可在15分钟内用肉眼直观检测到。未观察到与VP的交叉反应。此外，LFIA的灵敏度和特异性分别为94.0%和98.0%。所开发的试纸条可能会改变AHPND早期和原位诊断的局面。
