Abstract
Loop-mediated isothermal amplification (LAMP ), a DNA amplification method, has been applied to the detection of various pathogenic organisms because it features rapid reaction time, accuracy, cost-effect and results that can be determined with the naked eye. The most advantage of this method is that DNA amplification occurs at a constant temperature , usually between 60 and 65 °C, therefore sophisticated equipments are unnecessary. It can be easy that this method is employed in resource-limited laboratories and flied. We have designed a LAMP primer set to detect the kinetoplast minicircle DNA of Leishmania donovani . The LAMP was sensitive and could detect 1 fg of L. donovani DNA, and was not cross-reacted with DNA of 5 other Leishmania species, Plasmodium falciparum and human. The LAMP sensitivity and specificity is equal to nested PCR in DNA samples form patients with visceral leishmaniasis (VL). Therefore, the LAMP can be a better alternative to nested PCR, especially under field conditions, and will be a powerful tool for VL mass screening in combination with urine ELISA.
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Acknowledgments
Research underlying this chapter, conducted by the authors, was partially supported by JST/JICA, SATREPS, and also by a Grant-in-Aid for Scientific Research (B) No. 18406013 from the Japan Society for the Promotion of Science.
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Takagi, H., Itoh, M., Noiri, E. (2016). Loop-Mediated Isothermal Amplification (LAMP ): Molecular Diagnosis for the Field Survey of Visceral Leishmaniasis. In: Noiri, E., Jha, T. (eds) Kala Azar in South Asia. Springer, Cham. https://doi.org/10.1007/978-3-319-47101-3_14
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