Rapid diagnosis of Leishmaniasis by fluorogenic polymerase chain reaction

G. Wortmann; C. Sweeney; H. S. Houng; N. Aronson; J. Stiteler; J. Jackson; C. Ockenhouse

doi:10.4269/ajtmh.2001.65.583

Rapid diagnosis of Leishmaniasis by fluorogenic polymerase chain reaction

G. Wortmann^*, C. Sweeney, H. S. Houng, N. Aronson, J. Stiteler, J. Jackson, C. Ockenhouse

^*Corresponding author for this work

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Abstract

A fluorescent DNA probe (LEIS.P1) specific for a conserved region of the small-subunit ribosomal RNA gene of Leishmania and a pair of flanking primers (LEIS.U1 and LEIS.L1) were designed for use in a fluorogenic polymerase chain reaction. Optimal assay conditions with zero background were established to detect low levels of Leishmania from clinical samples. By use of this assay, we amplified DNA from 27 strains of cultured Leishmania (both Old and New World strains) and selectively amplified Leishmania DNA from 12 paraffin-embedded human biopsy samples and 3 fresh human skin biopsy specimens. For the fresh human tissue biopsies, the turnaround time from biopsy to test result was < 24 hr. No amplification was detected in negative control samples (including the kinetoplastid protozoa Trypanosoma rangelli and Crithidia fasiculata). This assay provides a specific and rapid diagnostic modality to detect infection with Leishmania.

Original language	English
Pages (from-to)	583-587
Number of pages	5
Journal	American Journal of Tropical Medicine and Hygiene
Volume	65
Issue number	5
DOIs	https://doi.org/10.4269/ajtmh.2001.65.583
State	Published - 2001
Externally published	Yes

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title = "Rapid diagnosis of Leishmaniasis by fluorogenic polymerase chain reaction",

abstract = "A fluorescent DNA probe (LEIS.P1) specific for a conserved region of the small-subunit ribosomal RNA gene of Leishmania and a pair of flanking primers (LEIS.U1 and LEIS.L1) were designed for use in a fluorogenic polymerase chain reaction. Optimal assay conditions with zero background were established to detect low levels of Leishmania from clinical samples. By use of this assay, we amplified DNA from 27 strains of cultured Leishmania (both Old and New World strains) and selectively amplified Leishmania DNA from 12 paraffin-embedded human biopsy samples and 3 fresh human skin biopsy specimens. For the fresh human tissue biopsies, the turnaround time from biopsy to test result was < 24 hr. No amplification was detected in negative control samples (including the kinetoplastid protozoa Trypanosoma rangelli and Crithidia fasiculata). This assay provides a specific and rapid diagnostic modality to detect infection with Leishmania.",

author = "G. Wortmann and C. Sweeney and Houng, {H. S.} and N. Aronson and J. Stiteler and J. Jackson and C. Ockenhouse",

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AU - Wortmann, G.

AU - Sweeney, C.

AU - Houng, H. S.

AU - Aronson, N.

AU - Stiteler, J.

AU - Jackson, J.

AU - Ockenhouse, C.

PY - 2001

Y1 - 2001

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AB - A fluorescent DNA probe (LEIS.P1) specific for a conserved region of the small-subunit ribosomal RNA gene of Leishmania and a pair of flanking primers (LEIS.U1 and LEIS.L1) were designed for use in a fluorogenic polymerase chain reaction. Optimal assay conditions with zero background were established to detect low levels of Leishmania from clinical samples. By use of this assay, we amplified DNA from 27 strains of cultured Leishmania (both Old and New World strains) and selectively amplified Leishmania DNA from 12 paraffin-embedded human biopsy samples and 3 fresh human skin biopsy specimens. For the fresh human tissue biopsies, the turnaround time from biopsy to test result was < 24 hr. No amplification was detected in negative control samples (including the kinetoplastid protozoa Trypanosoma rangelli and Crithidia fasiculata). This assay provides a specific and rapid diagnostic modality to detect infection with Leishmania.

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Rapid diagnosis of Leishmaniasis by fluorogenic polymerase chain reaction

Abstract

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