A neutralizing monoclonal antibody–based blocking ELISA to detect bovine herpesvirus 1 and vaccination efficacy

Liu, Wenxiao; Jiang, Hong; Duan, Jinglong; Jiang, Bo; Zhu, Rong; Cheng, Jing; Wang, Ping; Li, Yongqing

doi:10.1007/s00253-022-12308-z

Cited by 4 publications

(2 citation statements)

References 40 publications

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“…The specificity of mAbs was evaluated using western blotting and dot-blotting against BoHV-1, BVDV, BCoV, and BRV. ELISA additivity tests were utilised to identify whether mAbs recognised different BoHV-1 proteins and epitopes with reference to a previous study ( Liu et al, 2023 ).…”

Section: Methodsmentioning

confidence: 99%

“…The supernatants were discarded, and the virus-containing pellets were re-suspended in 50 mM Tris-HCl. Monoclonal antibodies (mAb) generated against BoHV-1 were prepared using purified BoHV-1 as the immunogen according to a previously described protocol (Liu et al, 2023). Briefly, mAbs were generated from ascites of BoHV-1-injected BALB/c mice with positive hybridoma cell lines and were purified using a HiTrap Protein A column (GE Healthcare, Memphis, TN, United States) according to the manufacturer's instructions.…”

Section: Cell Culture and Virus Purificationmentioning

confidence: 99%

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Development and evaluation of a time-resolved fluorescence labelled immunochromatographic strip assay for rapid and quantitative detection of bovine herpesvirus 1

Liu,

Zhang,

Cheng

et al. 2024

Front. Microbiol.

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Bovine herpes virus 1 (BoHV-1) causes a wide variety of diseases in wild and domestic cattle. The most widely used method for viral identification is real-time PCR, which can only be performed in laboratories using sophisticated instruments by expert personnel. Herein, we developed an ultrasensitive time-resolved fluorescence lateral flow immunochromatographic strip (ICS) assay for detecting BoHV-1 in bovine samples using a monoclonal antibody against BoHV-1 labelled with fluorescent microspheres, which can be applied in any setting. The intact process from sample collection to final result can be achieved in 15 min. The limit of detection of the assay for BoHV-1 was 102 TCID50/100 μL. The coincidence rate of the ICS method and real-time PCR recommended by the World Organization for Animal Health (WOAH) was 100% for negative, 92.30% for positive, and 95.42% for total, as evaluated by the detection of 131 clinical samples. This detection method was specifically targeted to BoHV-1, not exhibiting cross-reactivity with other bovine pathogens including BoHV-5. We developed an ICS assay equipped with a portable instrument that offers a sensitive and specific platform for the rapid and reliable detection of BoHV-1 in the field. The Point-of-Care test of BoHV-1 is suitable for the screening and surveillance of BoHV-1 in dairy herds.

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Section: Methodsmentioning

confidence: 99%

Section: Cell Culture and Virus Purificationmentioning

confidence: 99%