Imperial College London

DrJesusRodriguez Manzano

Faculty of MedicineDepartment of Infectious Disease

Non-Clinical Lecturer in Antimicrobial Resistance and Infect
 
 
 
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Contact

 

j.rodriguez-manzano

 
 
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Location

 

Electrical EngineeringSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Yu:2019:10.1016/j.jmoldx.2018.10.004,
author = {Yu, L-S and Rodriguez-Manzano, J and Malpartida-Cardenas, K and Sewell, T and Bader, O and Armstrong-James, D and Fisher, MC and Georgiou, P},
doi = {10.1016/j.jmoldx.2018.10.004},
journal = {Journal of Molecular Diagnostics},
pages = {286--295},
title = {Rapid and sensitive detection of azole-resistant Aspergillus fumigatus by tandem-repeat loop-mediated isothermal amplification},
url = {http://dx.doi.org/10.1016/j.jmoldx.2018.10.004},
volume = {21},
year = {2019}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Invasive human fungal infections caused by multi-azole resistant Aspergillus fumigatus are associated with increasing rates of mortality in susceptible patients. Current methods of diagnosing infections caused by multi-azole resistant A. fumigatus are, however, not well suited for use in clinical point-of-care testing or in the field. Loop-mediated isothermal amplification (LAMP) is a widely used method of nucleic acid amplification with rapid and easy-to-use features, making it suitable for use in different resource settings. Here, we developed a LAMP assay to detect a 34 bp tandem repeat, named TR34-LAMP. TR34 is a high-prevalence allele that, in conjunction with the L98H single nucleotide polymorphism, is associated with the occurrence of multi-azole resistance in A. fumigatus in the environment and in patients. This process was validated with both synthetic double stranded DNA and genomic DNA prepared from azole-resistant isolates of A. fumigatus. Use of our assay resulted in rapid and specific identification of the TR34 allele with high sensitivity, detecting down to 10 genomic copies per reaction within 25 minutes. Fluorescent and colorimetric detections were used for the analysis of 11 clinical isolates as cross validation. These results show that the TR34-LAMP assay has the potential to accelerate the screening of clinical and environmental A. fumigatus to provide a rapid and accurate diagnosis of azole resistance, which current methods struggle to achieve.
AU - Yu,L-S
AU - Rodriguez-Manzano,J
AU - Malpartida-Cardenas,K
AU - Sewell,T
AU - Bader,O
AU - Armstrong-James,D
AU - Fisher,MC
AU - Georgiou,P
DO - 10.1016/j.jmoldx.2018.10.004
EP - 295
PY - 2019///
SN - 1525-1578
SP - 286
TI - Rapid and sensitive detection of azole-resistant Aspergillus fumigatus by tandem-repeat loop-mediated isothermal amplification
T2 - Journal of Molecular Diagnostics
UR - http://dx.doi.org/10.1016/j.jmoldx.2018.10.004
UR - https://www.ncbi.nlm.nih.gov/pubmed/30529128
UR - http://hdl.handle.net/10044/1/65589
VL - 21
ER -