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Detection of white spot syndrome virus in seafood samples using a magnetosome-based impedimetric biosensor.
Archives of Virology ( IF 2.5 ) Pub Date : 2021-08-03 , DOI: 10.1007/s00705-021-05187-8
Sumana Sannigrahi 1 , Shiva Kumar Arumugasamy 2 , Jayaraman Mathiyarasu 2 , R Sudhakaran 3 , K Suthindhiran 4
Affiliation  

White spot syndrome virus (WSSV) is a significant threat to the aquaculture sector, causing mortality among crabs and shrimps. Currently available diagnostic tests for WSSV are not rapid or cost-effective, and a new detection method is therefore needed. This study demonstrates the development of a biosensor by functionalization of magnetosomes with VP28-specific antibodies to detect WSSV in seafood. The magnetosomes (1 and 2 mg/ml) were conjugated with VP28 antibody (0.025-10 ng/µl), as confirmed by spectroscopy. The magnetosome-antibody conjugate was used to detect the VP28 antigen. The binding of antigen to the magnetosome-antibody complex resulted in a change in absorbance. The magnetosome-antibody-antigen complex was then concentrated and brought near a screen-printed carbon electrode by applying an external magnetic field, and the antigen concentration was determined using impedance measurements. The VP28 antigen (0.025 ng/µl) bound more efficiently to the magnetosome-VP28 antibody complex (0.025 ng/µl) than to the VP28 antibody (0.1 ng/µl) alone. The same assay was repeated to detect the VP28 antigen (0.01 ng/µl) in WSSV-infected seafood samples using the magnetosome-VP28 antibody complex (0.025 ng/µl). The WSSV in the seafood sample was also drawn toward the electrode due to the action of magnetosomes controlled by the external magnetic field and detected using impedance measurement. The presence of WSSV in seafood samples was verified by Western blot and RT-PCR. Cross-reactivity assays with other viruses confirmed the specificity of the magnetosome-based biosensor. The results indicate that the use of the magnetosome-based biosensor is a sensitive, specific, and rapid way to detect WSSV in seafood samples.

中文翻译:

使用基于磁小体的阻抗生物传感器检测海鲜样品中的白斑综合征病毒。

白斑综合症病毒(WSSV)对水产养殖业构成重大威胁,导致螃蟹和虾死亡。目前可用的 WSSV 诊断测试既不快速也不划算,因此需要一种新的检测方法。这项研究证明了通过使用 VP28 特异性抗体对磁小体进行功能化来检测海鲜中的 WSSV 来开发生物传感器。磁小体(1 和 2 毫克/毫升)与 VP28 抗体(0.025-10 纳克/微升)结合,如光谱学所证实。磁小体-抗体偶联物用于检测VP28抗原。抗原与磁小体-抗体复合物的结合导致吸光度发生变化。然后通过施加外部磁场将磁小体-抗体-抗原复合物浓缩并使其靠近丝网印刷的碳电极,并且使用阻抗测量确定抗原浓度。与单独的 VP28 抗体 (0.1 ng/µl) 相比,VP28 抗原 (0.025 ng/µl) 与磁小体-VP28 抗体复合物 (0.025 ng/µl) 的结合效率更高。使用磁小体-VP28 抗体复合物 (0.025 ng/µl) 重复相同的测定以检测 WSSV 感染的海鲜样品中的 VP28 抗原 (0.01 ng/µl)。由于磁小体的作用受外部磁场控制并使用阻抗测量进行检测,海鲜样品中的 WSSV 也被拉向电极。通过蛋白质印迹和 RT-PCR 验证了海鲜样品中 WSSV 的存在。与其他病毒的交叉反应试验证实了基于磁小体的生物传感器的特异性。结果表明,使用基于磁小体的生物传感器是一种灵敏、特异、
更新日期:2021-08-03
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