TITLE:
The Development of a Loop-Mediated Isothermal Amplification (LAMP) Procedure for Plague Diagnostic
AUTHORS:
Mariana de Lira Nunes, Carina Lucena Mendes-Marques, Alzira Maria Paiva de Almeida, Nilma Cintra Leal
KEYWORDS:
Plague, Yersinia pestis, Diagnosis Tests, Loop-Mediated Isothermal Amplification (LAMP)
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.5 No.16,
November
25,
2014
ABSTRACT: Plague caused by Yersinia pestis is one of the infectious
diseases subject to the International Health Regulations (IHR). Permanent
monitoring of the focal plague areas is mandatory in order to enable prompt
control measures to prevent the spread of the disease. Therefore, the availability
of efficient diagnosis tests is of paramount importance. Here, we describe a
loop-mediated isothermal amplification (LAMP)-based procedure for rapid Y. pestis detection. We constructed a
set of LAMP primers, which were used in assays to establish the reaction conditions
that would lead to the quick visualization of the results by evaluating the
test tube with the naked eye. The primers were specifically designed to target
the caf1 gene located on pFra/Tox (pMT), a prototypical plasmid of Y. pestis. The LAMP procedure was performed
at 65°C for 45 min in a water bath and allowed for the detection of at least 10
pg of bacterial DNA. Due to its simplicity, specificity, sensitivity and
rapidity, the LAMP technique is an additional tool that may be implemented in
routine plague diagnoses, especially in emergencies.