RPB0528

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Japanese Encephalitis Virus Japanese encephalitis virus,Japanese encephalitis (JE) virus,Japanese encephalitis virus JE,Japanese encephalitis virus JEV 11072 Amarillovirales Flaviviridae Orthoflavivirus Orthoflavivirus japonicum Viruses

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM） GC Content（%） Predicted Melting Temperature(℃） Molecular Weight(g/moles) Positions in GenBank accession number
JEV-F1 GAACTCATCATTCCGCATACCATAGCCGGAC 31 10 μM 51.61 64.22 9409.18 \
JEV-R1 AGTGGTTCTGACCGAACGGCCTCTCTTGCGA 31 10 μM 58.06 70.04 9503.2 \
JEV-P FAM - TCGCAAGAGGTGGACGGCCA - BHQ 24 10 μM 60.42 66.49 7416.21 \

Gene Description

Target Gene GenBank ID
JEV NS1 \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
This study successfully developed a portable, dual-read, one-pot RPA-EsCas13d detection system that offers superior sensitivity and specificity for the rapid detection of JEV. RPA-CRISPR\EsCas13d Primer Premier 5.0 30 min 37 °C CRISPR\EsCas13d 8 × 10¹copies \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 Rapid, sensitive, and visual detection of swine Japanese encephalitis virus with a one-pot RPA-CRISPR\EsCas13d-based dual readout portable platform Dong You,Tong Xu,Bing-Zhou Huang,Ling Zhu,Fang Wu,Li-Shuang Deng,Zhe-Yan Liu,Jia-Qi Duan,Yuan-Meng Wang,Liang-Peng Ge,Zuo-Hua Liu,Jing Sun,Xiu Zeng,Li-Qiao Lang,Yuan-Cheng Zhou,Di-Shi Chen,Si-Yuan Lai,Yan-Ru Ai,Jian-Bo Huang,Zhi-Wen Xu International journal of biological macromolecules 39059534 10.1016/j.ijbiomac.2024.134151

Rapid, sensitive, and visual detection of swine Japanese encephalitis virus with a one-pot RPA-CRISPR\EsCas13d-based dual readout portable platform

Author(s):

Dong You,Tong Xu,Bing-Zhou Huang,Ling Zhu,Fang Wu,Li-Shuang Deng,Zhe-Yan Liu,Jia-Qi Duan,Yuan-Meng Wang,Liang-Peng Ge,Zuo-Hua Liu,Jing Sun,Xiu Zeng,Li-Qiao Lang,Yuan-Cheng Zhou,Di-Shi Chen,Si-Yuan Lai,Yan-Ru Ai,Jian-Bo Huang,Zhi-Wen Xu

Journal:

International journal of biological macromolecules

Year:

2024

Abstract:

Japanese encephalitis (JE), a mosquito-borne zoonotic disease caused by the Japanese encephalitis virus (JEV), poses a serious threat to global public health. The low viremia levels typical in JEV infections make RNA detection challenging, necessitating early and rapid diagnostic methods for effective control and prevention. This study introduces a novel one-pot detection method that combines recombinant enzyme polymerase isothermal amplification (RPA) with CRISPR/EsCas13d targeting, providing visual fluorescence and lateral flow assay (LFA) results. Our portable one-pot RPA-EsCas13d platform can detect as few as two copies of JEV nucleic acid within 1 h, without cross-reactivity with other pathogens. Validation against clinical samples showed 100 % concordance with real-time PCR results, underscoring the method's simplicity, sensitivity, and specificity. This efficacy confirms the platform's suitability as a novel point-of-care testing (POCT) solution for detecting and monitoring the JE virus in clinical and vector samples, especially valuable in remote and resource-limited settings.