RPB0183

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Mycobacterium tuberculosis Mycobacterium tuberculosis, Bacterium tuberculosis, Bacillus tuberculosis, Mycobacterium tuberculosis variant tuberculosis 1773 Corynebacteriales Mycobacteriaceae Mycobacterium Mycobacterium tuberculosis 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
F CCAAGCTGCGCCAGGGCAGCTATTTCCCGGAC 32 10 mM 65.6 73.84 9771.37 \
R Biotin-TTGGCCATGATCGACACTTGCGACTTGGA 29 10 mM 51.7 65.92 8908.83 \
P FITC-GAACGCGCACTGACCAGCGTGGTGGCGACCTG(H)TACCTGCTGGGAGTATC-P 49 10 mM 63.3 77.99 15139.82 \

Gene Description

Target Gene GenBank ID
IS1081 NC_000962.3

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
\ LF-RPA combined instruction manual of TwistAmp® DNA amplification kits (TwistDx Ltd., UK) 20 39 \ 12.5fg 100 97.92

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2017 Rapid and visual detection of Mycobacterium tuberculosis complex using recombinase polymerase amplification combined with lateral flow strips Qinglin Ma , Houming Liu , Feidi Ye , Guangxin Xiang , Wanshui Shan , Wanli Xing Molecular and Cellular Probes 28842221 10.1016/j.mcp.2017.08.004

Rapid and visual detection of Mycobacterium tuberculosis complex using recombinase polymerase amplification combined with lateral flow strips

Author(s):

Qinglin Ma , Houming Liu , Feidi Ye , Guangxin Xiang , Wanshui Shan , Wanli Xing

Journal:

Molecular and Cellular Probes

Year:

2017

Abstract:

To definitively diagnose active pulmonary Tuberculosis (TB), Mycobacterium tuberculosis complex (MTBC) bacilli must be identified within clinical specimens from patients. In this study, we introduced a rapid and visual detection method of MTBC using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strips. The LF-RPA assay, read results with naked eyes, could detect as few as 5 genome copies of M. tuberculosis H37Rv (ATCC 27294) per reaction and had no cross-reactions with other control bacteria even using excessive amount of template DNA. The system could work well at a broad range of temperature 25-45 °C and reach detectable level even within 5 min. When testing a total of 137 clinical specimens, the sensitivity and specificity of the LF-RPA assay were 100% (95% CI: 95.94%-100%) and 97.92% (95% CI: 88.93%-99.95%), respectively, compared to culture identification method. Therefore, the LF-RPA system we have demonstrated is a rapid, simple, robust method for MTBC detection which, subject to the availability of a suitable sample extraction method, has the potentiality to diagnose TB at the point-of-care testing.