RPB0229

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
RSV A/B Respiratory syncytial virus, Respiratory syncytial virus RS, respiratory syncytial virus RS virus, Respiratory syncytial virus RSV 12814 Mo\gavirales Pneumoviridae Orthopneumovirus Bovine orthopneumovirus 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
RSV-F2 CAGGATTGTTTATGAATGCCTATGGTKCAGG 31 600 nM 43.55 60.13 9608.79 \
RSV-R2 GGTGTACCTCTRTAYTCTCCCATTATGCCTA 31 600 nM 45.16 60.2 9387.65 \

Gene Description

Target Gene GenBank ID
N protein gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
detecting RSV and ADV is rapid, sensitive, and specific. RPA-CRISPR\Cas12a primer premier 5 20 min 40°C CRISPR\Cas detection system 10*4 copies 0.981 1

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 detecting RSV and ADV is rapid, sensitive, and specific. Gao Hongdan,Du Yao,Chai Qiang,Huang Meng,Liu Xiaorong,Xing Zhihao,Ma Dongli Journal Of International Medical Research 38230675 10.1177/03000605231223083

detecting RSV and ADV is rapid, sensitive, and specific.

Author(s):

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

Journal:

Journal Of International Medical Research

Year:

2024

Abstract:

Objective: Respiratory syncytial virus (RSV) and respiratory adenovirus (ADV) are two common pathogens that cause acute respiratory tract infections in children. We aimed to develop a rapid method for detecting both pathogens simultaneously. Methods: The recombinase polymerase isothermal amplification (RPA) method was combined with the CRISPR/Cas detection system. The assay's specificity and sensitivity were explored by designing RPA primers and CRISPR RNAs (crRNAs) through multi-sequence comparisons, optimizing the reaction conditions, and using a fluorescent reading device. The consistency of the test results of 160 clinical pharyngeal swab samples was studied using quantitative polymerase chain reaction (qPCR) results as a comparative control. Results: RSV and ADV could be detected at levels as low as 104 copies/mL and 103 copies/mL, respectively, within 50 minutes with no cross-reactivity with other similar pathogens. For the clinical samples, compared with the qPCR method, the sensitivities for RSV and ADV were 98.1% and 91.4%, respectively, and the detection specificities were both 100%. The Kappa values were greater than 0.95, suggesting a high degree of consistency. Conclusion: This method for detecting RSV and ADV is rapid, sensitive, and specific. It can accurately detect mixed infections in a timely manner, making it suitable for use in areas with scarce healthcare resources.