RPB0236

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
HAdV Human adenovirus 1907210 Rowavirales Adenoviridae Mastadenovirus Human adenovirus 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
F CTTATCCACCACATGGGAGCCAGGAGTTCC 30 2 μM 56.67 66.32 9152 \
R BIOTIN-TGCTCATGAACATGGCCAGATCGCACATTTCAG 33 2 μM 48.48 65.57 10097.62 \
P FAM-ACTGTCTCTGTCCGAGGTCCTGACGGTTCC[THF]TCCCATTAGTAACAAC-C3-SPACER 46 2 μM 52.17 70.74 14012.12 \

Gene Description

Target Gene GenBank ID
HAdV B 1A（E1A）gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
rapid nucleic acid detection of HAdV B using recombinase polymerase amplification assay (RPA) RPA Oligo5 30 min 40°C \ 10 copies 0.878 100 %

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 Rapid detection of human adenovirus subgroup B using recombinase polymerase amplification assay Yongzhe Zhu,Binghui Xia,Haizhou Xu,Zengxin Liu,Ru Wang,Qingqing Cai,Ping Zhao,Zhongtian Qi Virus Genes 38175387 10.1007/s11262-023-02044-5

Rapid detection of human adenovirus subgroup B using recombinase polymerase amplification assay

Author(s):

Yongzhe Zhu,Binghui Xia,Haizhou Xu,Zengxin Liu,Ru Wang,Qingqing Cai,Ping Zhao,Zhongtian Qi

Journal:

Virus Genes

Year:

2024

Abstract:

Human adenovirus subgroup B (HAdV B) is one of the major pathogens of human respiratory virus infections, which has considerable transmission and morbidity in a variety of populations. Therefore, rapid and specific detection of HAdV B in clinical samples is essential for diagnosis. This study aimed to develop a product for rapid nucleic acid detection of HAdV B using recombinase polymerase amplification assay (RPA) and validate the performance of this method by using clinical samples. Results showed that this method achieved a lower limit of detection (LOD) of 10 copies/μL and had no cross-reactivity with other adenovirus subgroups or respiratory pathogens. In addition to high sensitivity, it can be completed within 30 min at 40 °C. There is no need to perform nucleic acid extraction on clinical samples. Taking qPCR as the gold standard, the RPA assay possessed a high concordance (Cohen's kappa, 0.896; 95% CI 0.808-0.984; P < 0.001), with a sensitivity of 87.80% and a specificity of 100.00%. The RPA assay developed in this study provided a simple and highly specific method, making it an important tool for rapid adenovirus nucleic acid detection and facilitating large-scale population screening in resource-limited settings.