Generation and Characterization of Monoclonal Antibodies Against Red-Spotted Grouper Nervous Necrosis Virus (RGNNV)

Qin, Yinghui; Liu, Jiaoyun; Lu, Yuanan; Liu, Xueqin

doi:10.1007/s11802-022-4895-4

Cited by 1 publication

(2 citation statements)

References 20 publications

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“…For instance, while we obtained 40 positive monoclonal hybridomas recognizing rMCP through ELISA screening, only two of them were able to recognize native MCP in western blot and IFA using LMBV-infected EPC cells. The 5% positive rate was lower than that in our previous study (Qin et al, 2022). The low positive rate could be attributed to the recombinant MCP expressed in E.coil lacking similar posttranslational modifications as in fish or eukaryotes, which could lead to changes in epitopes related to the native MCP of LMBV (Zhao et al, 2007;Macek et al, 2019).…”

Section: Discussioncontrasting

confidence: 71%

“…The fused cells were then resuspended in 1640 medium (Biosharp, China) containing 1% HAT (Sigma, USA) and seeded into 96-well culture plates (100 mL/well). After 14 days, the supernatant from those wells containing hybridomas was collected and screened using indirect enzyme-linked immunosorbent assay (ELISA), western blot, and indirect immunofluorescence assay (IFA) as per a previous method (Qin et al, 2022). The positive hybridomas were subcloned using the limited dilution method until all monoclonal cells' supernatant reacted with MCP.…”

Section: Preparation and Characterization Of Monoclonal Antibody Agai...mentioning

confidence: 99%

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Isolation, identification, and monoclonal antibody development of largemouth bass virus

Qin,

Liu,

Mao

et al. 2024

Front. Mar. Sci.

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Largemouth bass virus (LMBV) poses a significant threat to largemouth bass farming, leading to substantial economic losses. In December 2022, massive largemouth bass juveniles died at a fish farm in the city of Xinxiang, China. Through a series of experiments, we conclusively identified LMBV as the causative pathogen. The affected fish displayed anorexia, lethargy, and hemorrhage at the pectoral and caudal fin base. No parasites or pathogenic bacteria were detected on the body surface or gills, or isolated from the diseased fish. Severe hemorrhage, lymphocyte infiltration, and extensive necrosis were observed in the liver, spleen, intestine, and stomach of the moribund fish. The tissue homogenate from the diseased fish induced epithelioma papulosum cyprini cells (EPC) cell death, while no such effects were observed in grouper spleen (GS) cells. Sequence similarity analysis of the major capsid protein (MCP) indicated the virus shared 100% similarity with the LMBV-FS2021 strain, placing it within the Ranavirus genus. Transmission electron microscopy (TEM) observations revealed plenty of hexagonal virions accumulated in the cytoplasm of infected EPC cells. Artificial infection demonstrated that LMBV-XX01 was highly fatal to Micropterus salmoides juveniles, with an LD50 of 103.081 TCID50/fish. RT-qPCR detection confirmed that LMBV appeared in all sampled tissues of the challenged largemouth bass, with significantly higher viral loads detected in the liver and heart compared to other tissues. Additionally, we successfully obtained a highly purified recombinant MCP of LMBV and developed two strains of monoclonal antibodies targeting MCP of LMBV-XX01. Overall, our findings provide valuable materials and insights for the design of prevention strategies and the development of detection methods for LMBV.

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Section: Discussioncontrasting

confidence: 71%

Section: Preparation and Characterization Of Monoclonal Antibody Agai...mentioning

confidence: 99%

Isolation, identification, and monoclonal antibody development of largemouth bass virus

Qin,

Liu,

Mao

et al. 2024

Front. Mar. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

Generation and Characterization of Monoclonal Antibodies Against Red-Spotted Grouper Nervous Necrosis Virus (RGNNV)

Cited by 1 publication

References 20 publications

Isolation, identification, and monoclonal antibody development of largemouth bass virus

Isolation, identification, and monoclonal antibody development of largemouth bass virus

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