Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients

Juan David Ramírez; Giovanny Herrera; Carlos Muskus; Claudia Mendez; María Clara Duque; Robert Butcher

doi:10.1016/j.ijid.2018.10.029

Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients

Juan David Ramírez, Giovanny Herrera, Carlos Muskus, Claudia Mendez, María Clara Duque, Robert Butcher

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Abstract

Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform.

Original language	English (US)
Pages (from-to)	1-3
Number of pages	3
Journal	International Journal of Infectious Diseases
Volume	79
DOIs	https://doi.org/10.1016/j.ijid.2018.10.029
State	Published - Feb 1 2019

All Science Journal Classification (ASJC) codes

Microbiology (medical)
Infectious Diseases

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10.1016/j.ijid.2018.10.029

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abstract = "Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform.",

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AU - Herrera, Giovanny

AU - Muskus, Carlos

AU - Mendez, Claudia

AU - Duque, María Clara

AU - Butcher, Robert

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AB - Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform.

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Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients

Abstract

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