TY - JOUR
T1 - Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection
AU - Laurent, Thierry
AU - Van der Auwera, Gert
AU - Hide, Mallorie
AU - Mertens, Pascal
AU - Quispe-Tintaya, Wilber
AU - Deborggraeve, Stijn
AU - De Doncker, Simonne
AU - Leclipteux, Thierry
AU - Bañuls, Anne Laure
AU - Büscher, Philippe
AU - Dujardin, Jean Claude
N1 - Funding Information:
The authors are grateful to J.P. Dedet (University of Montpellier, France), G. Schönian (Institut für Mikrobiologie und Hygiene, Berlin, Germany), and I. Mauricio (London School of Hygiene and Tropical Medicine, London, UK) for sharing reference strains and DNA. We also acknowledge S. Rijal (B.P. Koirala Institute of Health Sciences, Dharan, Nepal), C. Cañavate (Instituto de Salud Carlos III, Majadahonda, Spain), and H. Louzir (Institut Pasteur de Tunis, Tunis-Belvédère, Tunisia) for providing clinical samples and cryostabilates. This study received financial support from the European Union's 5th and 6th framework programs “Leishmania Genotyping” (QLK2-CT-2001-01810) and “Tryleidiag” (INCO-CT-2005-015379).
PY - 2009/2
Y1 - 2009/2
N2 - 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.
AB - 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.
KW - Cysteine protease B
KW - Differential diagnosis
KW - Dipstick
KW - Leishmaniasis
KW - Oligochromatography
KW - Trypanosomatidae
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U2 - 10.1016/j.diagmicrobio.2008.10.015
DO - 10.1016/j.diagmicrobio.2008.10.015
M3 - Article
C2 - 19097841
AN - SCOPUS:58149496097
SN - 0732-8893
VL - 63
SP - 173
EP - 181
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 2
ER -