The Development of a Loop-Mediated Isothermal Amplification (LAMP) Procedure for Plague Diagnostic

• Published in: American journal of analytical chemistry 2014-11, Vol.5 (16), p.1069-1077
• Main Authors: de Lira Nunes, Mariana, Mendes-Marques, Carina Lucena, de Almeida, Alzira Maria Paiva, Leal, Nilma Cintra
• Format: Article
• Language: English
• Subjects: Yersinia pestis
• ISSN: 2156-8251; 2156-8278
• DOI: 10.4236/ajac.2014.516114

Plague caused by Yersinia pestis is one of the infectious diseases subject to the International Health Regulations. 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. In this article, the researchers describe a loop-mediated isothermal amplification (LAMP)-based procedure for rapid Y. pestis detection. They 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, a prototypical plasmid of Y. pestis. The LAMP procedure was performed at 65 degrees Celsius for 45 min in a water bath and allowed for the detection of at least 10 pg of bacterial DNA.