Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection

Thierry Laurent, Gert Van der Auwera, Mallorie Hide, Pascal Mertens, Wilber Quispe-Tintaya, Stijn Deborggraeve, Simonne De Doncker, Thierry Leclipteux, Anne Laure Bañuls, Philippe Büscher, Jean Claude Dujardin

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We used the cysteine proteinase B (cpb) gene family of the trypanosomatid genus Leishmania as a target to develop rapid, specific, and easy-to-use polymerase chain reaction (PCR) tests to discriminate Leishmania infantum, Leishmania donovani, Leishmania tropica, Leishmania aethiopica, and Leishmania major. Identification of all 5 Old World species and validation of intraspecies variability are features lacking in other species-specific PCRs. Amplicon analysis was done on agarose gels and was further simplified by using an oligochromatography dipstick to detect L. infantum and L. donovani products. Because the analytical sensitivity is lower than that of certain other species- and genus-specific PCRs, our assays are especially valuable for use on cultured isolates or directly on cryostabilates. As such, they can be implemented by research and health centers having access to culturing, DNA isolation, and PCR.

Original languageEnglish (US)
Pages (from-to)173-181
Number of pages9
JournalDiagnostic Microbiology and Infectious Disease
Volume63
Issue number2
DOIs
StatePublished - Feb 2009

Fingerprint

yeast proteinase B
Cysteine Proteases
Leishmania
Leishmania infantum
Leishmania donovani
Polymerase Chain Reaction
Genes
Leishmania tropica
Leishmania major
DNA-Directed DNA Polymerase
Sepharose
Gels
Health
Research

Keywords

  • Cysteine protease B
  • Differential diagnosis
  • Dipstick
  • Leishmaniasis
  • Oligochromatography
  • Trypanosomatidae

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

Cite this

Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection. / Laurent, Thierry; Van der Auwera, Gert; Hide, Mallorie; Mertens, Pascal; Quispe-Tintaya, Wilber; Deborggraeve, Stijn; De Doncker, Simonne; Leclipteux, Thierry; Bañuls, Anne Laure; Büscher, Philippe; Dujardin, Jean Claude.

In: Diagnostic Microbiology and Infectious Disease, Vol. 63, No. 2, 02.2009, p. 173-181.

Research output: Contribution to journalArticle

Laurent, T, Van der Auwera, G, Hide, M, Mertens, P, Quispe-Tintaya, W, Deborggraeve, S, De Doncker, S, Leclipteux, T, Bañuls, AL, Büscher, P & Dujardin, JC 2009, 'Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection', Diagnostic Microbiology and Infectious Disease, vol. 63, no. 2, pp. 173-181. https://doi.org/10.1016/j.diagmicrobio.2008.10.015
Laurent, Thierry ; Van der Auwera, Gert ; Hide, Mallorie ; Mertens, Pascal ; Quispe-Tintaya, Wilber ; Deborggraeve, Stijn ; De Doncker, Simonne ; Leclipteux, Thierry ; Bañuls, Anne Laure ; Büscher, Philippe ; Dujardin, Jean Claude. / Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection. In: Diagnostic Microbiology and Infectious Disease. 2009 ; Vol. 63, No. 2. pp. 173-181.
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