Epidemiological investigation of swine Japanese encephalitis virus based on RT-RAA detection method

Nie, Mincai; Zhou, Yuancheng; Li, Fengqin; Deng, Huidan; Zhao, Mingyang; Huang, Yao; Jiang, Chaoyuan; Sun, Xiangang; Xu, Zhiwen; Zhu, Ling

doi:10.1038/s41598-022-13604-4

Cited by 8 publications

(6 citation statements)

References 27 publications

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“…Globally, approximately 68,000 cases of Japanese encephalitis virus infection occur annually, with children under 14 years old accounting for 75% of the total number. In animals, pigs infected with JEV usually suffer from reproductive disorders causing huge economic losses to the pig industry [ 49 , 50 ]. And, no effective antiviral drugs for JEV are currently available [ 51 , 52 ].…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Knockout of the HuR Gene in U251 Cell Inhibits Japanese Encephalitis Virus Replication

Luo,

Cao,

Zhao

2024

Microorganisms

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Human antigen R (HuR) is an RNA-binding protein that regulates the post-transcriptional reaction of its target mRNAs. HuR is a critical factor in cancer development and has been identified as a potential target in many cancer models. It participates in the viral life cycle by binding to viral RNAs. In prior work, we used CRISPR/Cas9 screening to identify HuR as a prospective host factor facilitating Japanese encephalitis virus (JEV) infection. The HuR gene was successfully knocked out in U251 cell lines using the CRISPR/Cas9 gene-editing system, with no significant difference in cell growth between U251-WT and U251-HuR-KO2 cells. Here, we experimentally demonstrate for the first time that the knockout of the HuR gene inhibits the replication ability of JEV in U251 cell lines. These results play an essential role in regulating the replication level of JEV and providing new insights into virus–host interactions and potential antiviral strategies. It also offers a platform for investigating the function of HuR in the life cycle of flaviviruses.

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Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Knockout of the HuR Gene in U251 Cell Inhibits Japanese Encephalitis Virus Replication

Luo,

Cao,

Zhao

2024

Microorganisms

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“…Overall, PHEV, PRV, CSFV, and JEV are still popular in many countries around the world, and cause different degrees of damages to the pig industry (5,6,13,14,20,21,28,29,57). In China, some provinces have recently reported the prevalence of PHEV, PRV, CSFV, and JEV (41,47,51,(58)(59)(60)(61)(62)(63)(64)(65). The nasal cavity or throat swabs from 1-to 3-week-old piglets in Jilin province in 2010 showed a positivity rate of 44.23% (207/468) for PHEV (58).…”

Section: Discussionmentioning

confidence: 99%

Development of a quadruplex real-time quantitative RT-PCR for detection and differentiation of PHEV, PRV, CSFV, and JEV

Hu,

Feng,

Shi

et al. 2023

Front. Vet. Sci.

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Porcine hemagglutinating encephalomyelitis virus (PHEV), porcine pseudorabies virus (PRV), classical swine fever virus (CSFV), and Japanese encephalitis virus (JEV) cause similar neurological symptoms in the infected pigs, and their differential diagnosis depends on laboratory testing. Four pairs of specific primers and probes were designed targeting the PHEV N gene, PRV gB gene, CSFV 5′ untranslated region (5’UTR), and JEV NS1 gene, respectively, and a quadruplex real-time quantitative RT-PCR (qRT-PCR) was developed to detect and differentiate PHEV, PRV, CSFV, and JEV. The assay showed high sensitivity, with the limit of detection (LOD) of 1.5 × 101 copies/μL for each pathogen. The assay specifically detected only PHEV, PRV, CSFV, and JEV, without cross-reaction with other swine viruses. The coefficients of variation (CVs) of the intra-assay and the inter-assay were less than 1.84%, with great repeatability. A total of 1,977 clinical samples, including tissue samples, and whole blood samples collected from Guangxi province in China, were tested by the developed quadruplex qRT-PCR, and the positivity rates of PHEV, PRV, CSFV, and JEV were 1.57% (31/1,977), 0.35% (7/1,977), 1.06% (21/1,977), and 0.10% (2/1,977), respectively. These 1,977 samples were also tested by the previously reported qRT-PCR assays, and the coincidence rates of these methods were more than 99.90%. The developed assay is demonstrated to be rapid, sensitive, and accurate for detection and differentiation of PHEV, PRV, CSFV, and JEV.

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“…This approach involves the introduction of recombinase and single-stranded binding protein into the amplification system. RT-RAA achieves rapid amplification at 39°C, with results obtainable within minutes ( Nie et al, 2022 ). A key advantage of RT-RAA is its capacity to directly utilize RNA as a template for pathogen detection.…”

Section: Isothermal Amplification Techniquesmentioning

confidence: 99%

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

Mao,

Xu,

Luo

et al. 2023

Front. Bioeng. Biotechnol.

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In the realm of pathogen detection, isothermal amplification technology has emerged as a swift, precise, and sensitive alternative to conventional PCR. This paper explores the fundamental principles of recombinase polymerase amplification (RPA) and recombinase-aid amplification (RAA) and reviews the current status of integrating the CRISPR-Cas system with RPA/RAA techniques. Furthermore, this paper explores the confluence of isothermal amplification and CRISPR-Cas technology, providing a comprehensive review and enhancements of existing combined methodologies such as SHERLOCK and DETECTR. We investigate the practical applications of RPA/RAA in conjunction with CRISPR-Cas for pathogen detection, highlighting how this integrated approach significantly advances both research and clinical implementation in the field. This paper aims to provide readers with a concise understanding of the fusion of RPA/RAA and CRISPR-Cas technology, offering insights into their clinical utility, ongoing enhancements, and the promising prospects of this integrated approach in pathogen detection.

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Epidemiological investigation of swine Japanese encephalitis virus based on RT-RAA detection method

Cited by 8 publications

References 27 publications

CRISPR/Cas9-Mediated Knockout of the HuR Gene in U251 Cell Inhibits Japanese Encephalitis Virus Replication

CRISPR/Cas9-Mediated Knockout of the HuR Gene in U251 Cell Inhibits Japanese Encephalitis Virus Replication

Development of a quadruplex real-time quantitative RT-PCR for detection and differentiation of PHEV, PRV, CSFV, and JEV

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

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