RPB0097

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Haemophilus influenzae Haemophilus influenzae, Bacterium influenzae, Mycobacterium influenzae, Influenza-bacillus, Coccobacillus pfeifferi, Haemophilus meningitidis 727 Pasteurellales Pasteurellaceae Haemophilus Haemophilus influenzae Bacterium

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
bexA-F3 CAGGAAATGGTGCTGCTCAAACTTTTGGCGGTTAC 35 0.42 48.6 66.05 10802.06 \
bexA-R1B Biotin-TGTGAAACGAAAATGATAGAACGGTCTTTGC 31 0.42 38.7 58.9 9607.33 \
bexA-P FITC-CGGTTGAGTATGATTGTTATGTAATTGATGAG[THF]TGATTGTAGTAGGG-C3 spacer 46 0.12 37 61.59 14415.4 \

Gene Description

Target Gene GenBank ID
bexA \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
Detection of Capsulated and Non-Capsulated Haemophilus influenzae RPA-LFS Primer Premier 5.0 software 30 37 lateral flow strip (LFS) 100 CFU/reaction \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2022 Establishment and Clinical Application of a RPA-LFS Assay for Detection of Capsulated and Non-Capsulated Haemophilus influenzae Yan Wang,Aibo Liu,Mei Fu,Jingjing Guo,Lei Wang,Xiaohua Zuo,Fenfen Ma Frontiers in cellular and infection microbiology 35531333 10.3389/fcimb.2022.878813

Establishment and Clinical Application of a RPA-LFS Assay for Detection of Capsulated and Non-Capsulated Haemophilus influenzae

Author(s):

Yan Wang,Aibo Liu,Mei Fu,Jingjing Guo,Lei Wang,Xiaohua Zuo,Fenfen Ma

Journal:

Frontiers in cellular and infection microbiology

Year:

2022

Abstract:

A recombinase polymerase amplification-lateral flow strip assay was established for detection of the outer membrane protein P6 (omp6) and the capsule encoding gene bexA of Haemophilus influenzae and the detection limit, sensitivity, and specificity were determined. Specific primers and probes were designed based on the published nucleotide sequences of omp6 and bexA. The minimum detection limit was determined with standard strains and the practical applicability of the RPA-LFS assay was assessed by detection of 209 clinical samples. The results confirmed that the RPA-LFS assay was both specific and sensitive for the detection of capsulated and non-capsulated H. influenzae with a detection limit of 1 CFU/µL. The detection rate of the 209 clinical samples was 97.1%, while the detection rate of capsulated H. influenzae was 63.2%. The detection results were consistent with the traditional culture method and dual polymerase chain reaction (PCR), confirming the applicability of the RPA-LFS assay.