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Keywords = luciferase immunosorbent assay (LISA)

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13 pages, 2415 KiB  
Article
Development of a Luciferase Immunosorbent Assay for Detecting Crimean–Congo Hemorrhagic Fever Virus IgG Antibodies Based on Nucleoprotein
by Qi Chen, Yuting Fang, Ning Zhang and Chengsong Wan
Viruses 2025, 17(1), 32; https://doi.org/10.3390/v17010032 - 28 Dec 2024
Viewed by 1141
Abstract
Crimean–Congo hemorrhagic fever (CCHF) is a serious tick-borne disease with a wide geographical distribution. Classified as a level 4 biosecurity risk pathogen, CCHF can be transmitted cross-species due to its aerosol infectivity and ability to cause severe hemorrhagic fever outbreaks with high morbidity [...] Read more.
Crimean–Congo hemorrhagic fever (CCHF) is a serious tick-borne disease with a wide geographical distribution. Classified as a level 4 biosecurity risk pathogen, CCHF can be transmitted cross-species due to its aerosol infectivity and ability to cause severe hemorrhagic fever outbreaks with high morbidity and mortality. However, current methods for detecting anti-CCHFV antibodies are limited. This study aimed to develop a novel luciferase immunosorbent assay (LISA) for the detection of CCHFV-specific IgG antibodies. We designed specific antigenic fragments of the nucleoprotein and evaluated their sensitivity and specificity in detecting IgG in serum samples from mice and horses. In addition, we compared the efficacy of our LISA to a commercial enzyme-linked immunosorbent assay (ELISA). Our results demonstrated that the optimal antigen for detecting anti-CCHFV IgG was located within the stalk cut-off domain of the nucleoprotein. The LISA exhibited high specificity for serum samples from indicated species and significantly higher sensitivity (at least 128 times) compared with the commercial ELISA. The proposed CCHFV-LISA has the potential to facilitate serological diagnosis and epidemiological investigation of CCHFV in natural foci, providing valuable technical support for surveillance and early warning of this disease. Full article
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11 pages, 2191 KiB  
Article
A Luciferase Immunosorbent Assay Based on Attachment Glycoprotein for the Rapid and Easy Detection of Nipah Virus IgG Antibodies
by Xinyue Li, Yuting Fang, Xinyi Huang, Yongkun Zhao and Chengsong Wan
Microorganisms 2024, 12(5), 983; https://doi.org/10.3390/microorganisms12050983 - 14 May 2024
Cited by 2 | Viewed by 1826
Abstract
Nipah virus (NiV) is a virulent zoonotic disease whose natural host is the fruit bat (Pteropus medius), which can coexist with and transmit the virus. Due to its high pathogenicity, wide host range, and pandemic potential, establishing a sensitive, specific, and [...] Read more.
Nipah virus (NiV) is a virulent zoonotic disease whose natural host is the fruit bat (Pteropus medius), which can coexist with and transmit the virus. Due to its high pathogenicity, wide host range, and pandemic potential, establishing a sensitive, specific, and rapid diagnostic method for NiV is key to preventing and controlling its spread and any outbreaks. Here, we established a luciferase immunosorbent assay (LISA) based on the NiV attachment glycoprotein (G) to detect NiV-specific immunoglobulin G by expressing a fusion protein of nanoluciferase (NanoLuc) and the target antigen. Sensitivity analysis was performed and compared to an indirect enzyme-linked immunosorbent assay (ELISA), and specificity and cross-reactivity assessments were performed using NiV-positive horse serum and Ebola virus-, Crimean–Congo hemorrhagic fever virus-, and West Nile virus-positive horse sera. The optimal structural domain for NiV detection was located within amino acids 176–602 of the NiV G protein head domain. Moreover, the LISA showed at least fourfold more sensitivity than the indirect ELISA, and the cross-reactivity results suggested that the LISA had good specificity and was capable of detecting NiV-specific immunoglobulin G in both mouse and horse serum. In conclusion, the establishment of a rapid, simple NiV LISA using the G protein head domain provides a resource for NiV monitoring. Full article
(This article belongs to the Section Microbial Biotechnology)
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