RPB0235

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
HMPV Human metapneumovirus 162145 Mo\gavirales Pneumoviridae Metapneumovirus Human metapneumovirus Virus

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
HMPV-F1 ATGTCTCTTCAAGGGATTCACCTRAGTG 28 20μM 44.64 58.95 8586.64 \
HMPV-R1 ATYCCTATYTCTGCAGCATATTTGTAATC 29 20μM 34.48 54.33 8792.79 \

Gene Description

Target Gene GenBank ID
nucleoprotein (N) gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
A rapid, simple, sensitive and specific detection method of HMPV is presented. RT-RPA-CRISPR-Cas12a HMPV primers 30 min 37℃ CRISPR Cas12a system 1 copy/μL 0.9722 100 %

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 Rapid and one-tube detection of human metapneumovirus using the RT-RPA and CRISPR\Cas12a Yao Du,Xiaorong Liu,Hongdan Gao,Xiaoqian Liu,Meng Huang,Qiang Chai,Zhihao Xing,Tao Zhang,Dongli Ma Journal of Virological Methods 39038660 10.1016/j.jviromet.2024.115001

Rapid and one-tube detection of human metapneumovirus using the RT-RPA and CRISPR\Cas12a

Author(s):

Yao Du,Xiaorong Liu,Hongdan Gao,Xiaoqian Liu,Meng Huang,Qiang Chai,Zhihao Xing,Tao Zhang,Dongli Ma

Journal:

Journal of Virological Methods

Year:

2024

Abstract:

Human metapneumovirus (HMPV) is a common pathogen that can cause acute respiratory tract infections and is prevalent worldwide. There is yet no effective vaccine or specific treatment for HMPV. Early, rapid, and accurate detection is essential to treat the disease and control the spread of infection. In this study, we created the One-tube assay by combining Reverse Transcription-Recombinase Polymerase Amplification (RT-RPA) with the CRISPR/Cas12a system. By targeting the nucleoprotein (N) gene of HMPV to design specific primers and CRISPR RNAs (crRNAs), combining RT-RPA and CRISPR/Cas12a, established the One-tube assay. Meanwhile, the reaction conditions of the One-tube assay were optimized to achieve rapid and visual detection of HMPV. This assay could detect HMPV at 1 copy/μL in 30 min, without cross-reactivity with nine other respiratory pathogens. We validated the detection performance using clinical specimens and showed that the coincidence rate was 98.53 %，compared to the quantitative reverse-transcription polymerase chain reaction. The One-tube assay reduced the detection time and simplified the manual operation, while maintaining the detection performance and providing a new platform for HMPV detection.