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Comparison of LAMP and PCR for molecular mass screening of sand flies for Leishmania martiniquensis infection. | LitMetric

AI Article Synopsis

  • Leishmaniasis, caused by the Leishmania martiniquensis parasite, has been reported in various regions including Martinique and parts of Europe and the U.S., emphasizing the need for effective diagnostic tools to identify sand fly vectors for prevention.
  • The study aimed to validate two novel techniques, malachite green-LAMP (MG-LAMP) and PCR-mkDNA, for detecting L. martiniquensis in sand flies, comparing them to the established PCR-ITS1 method.
  • Results indicated that PCR-mkDNA was significantly more sensitive and specific than the other methods, successfully detecting the parasite in sand flies, but ultimately found no traces of Leishmania DNA in 380 samples from an affected area in

Article Abstract

Background: Leishmaniasis caused by Leishmania martiniquensis infection has been reported in human and domestic animals of Martinique Island, Germany, Switzerland, USA, Myanmar and Thailand. The peculiar clinical features of disseminated cutaneous and visceral forms co-existence render the urgent need of specific diagnostic tool to identify the natural sand fly vectors for effective prevention and control strategies. Loop-mediated isothermal amplification (LAMP) of 18S rRNA gene as well as polymerase chain reaction (PCR) of minicircle kinetoplast DNA gene (PCR-mkDNA) have never been applied to detect L. martiniquensis and L. siamensis in sand fly vectors.

Objective: The present study was aimed to validate malachite green-LAMP (MG-LAMP) and PCR-mkDNA techniques to detect L. martiniquensis in sand fly vectors, compared with the conventional PCR of internal transcribed spacer 1 (PCR-ITS1).

Methods: We compared the validity of LAMP of 18S rRNA gene and PCR-mkDNA, to PCR-ITS1 in simulation model of L. martiniquensis infection in Sergentomyia gemmea sand flies. Attributable to the sensitivity and specificity, PCR-mkDNA was consecutively applied to detect L. martiniquensis in 380 female sand fly individuals captured in the newly identified affected region of Lamphun Province, Thailand.

Findings And Main Conclusions: Results showed that PCR-mkDNA could detect at least one promastigote per sand fly, which was 10-time superior to LAMP and PCR-ITS1. In addition, PCR-mkDNA was more specific, able to differentiate L. martiniquensis from other viscerotropic Leishmania species, such as L. siamensis, L. (L.) donovani, and L. (L.) infantum. Consecutively, mass screening of L. martiniquensis in 380 female sand fly individuals by PCR-mkDNA was implemented in a new affected area of Thailand where a patient with leishmaniasis/HIV co-infection resides; however Leishmania DNA was undetected. In conclusion, PCR-mkDNA is a promising tool for molecular mass screening of L. martiniquensis infection in outbreak areas where several species of Leishmania and sand flies co-exist.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5293119PMC
http://dx.doi.org/10.1590/0074-02760160254DOI Listing

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