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{Malpartida-Cardenas:2018,
author = {Malpartida-Cardenas, K and Rodriguez-Manzano, J and Yu, L-S and Delves, M and Nguon, C and Chotivanich, K and Baum, J and Georgiou, P},
journal = {Analytical Chemistry},
pages = {11972--11980},
title = {Allele-specific isothermal amplification method using novel unmodified self-stabilizing competitive primers},
url = {http://hdl.handle.net/10044/1/64956},
volume = {90},
year = {2018}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Rapid and specific detection of single nucleotide polymorphisms (SNPs) related to drug resistance in infectious diseases is crucial for accurate prognostics, therapeutics and disease management at point-of-care. Here, we present a novel amplification method and provide universal guidelines for the detection of SNPs at isothermal conditions. This method, called USS-sbLAMP, consists of SNP-based loop-mediated isothermal amplification (sbLAMP) primers and unmodified self-stabilizing (USS) competitive primers that robustly delay or prevent unspecific amplification. Both sets of primers are incorporated into the same reaction mixture, but always targeting different alleles; one set specific to the wild type allele and the other to the mutant allele. The mechanism of action relies on thermodynamically favored hybridization of totally complementary primers, enabling allele-specific amplification. We successfully validate our method by detecting SNPs, C580Y and Y493H, in the Plasmodium falciparum kelch 13 gene that are responsible for resistance to artemisinin-based combination therapies currently used globally in the treatment of malaria. USS-sbLAMP primers can efficiently discriminate between SNPs with high sensitivity (limit of detection of 5 × 101 copies per reaction), efficiency, specificity and rapidness (<35 min) with the capability of quantitative measurements for point-of-care diagnosis, treatment guidance, and epidemiological reporting of drug-resistance.
AU - Malpartida-Cardenas,K
AU - Rodriguez-Manzano,J
AU - Yu,L-S
AU - Delves,M
AU - Nguon,C
AU - Chotivanich,K
AU - Baum,J
AU - Georgiou,P
EP - 11980
PY - 2018///
SN - 0003-2700
SP - 11972
TI - Allele-specific isothermal amplification method using novel unmodified self-stabilizing competitive primers
T2 - Analytical Chemistry
UR - http://hdl.handle.net/10044/1/64956
VL - 90
ER -