TY - JOUR
T1 - Single locus genotyping to track Leishmania donovani in the Indian subcontinent
T2 - application in Nepal
AU - Rai, Keshav
AU - Bhattarai, Narayan Raj
AU - Vanaerschot, Manu
AU - Imamura, Hideo
AU - Gebru, Gebreyohans
AU - Khanal, Basudha
AU - Rijal, Suman
AU - Boelaert, Marleen
AU - Pal, Chiranjib
AU - Karki, Prahlad
AU - Dujardin, Jean-Claude
AU - Van der Auwera, Gert
N1 - FTX; DOAJ
PY - 2017
Y1 - 2017
N2 - BACKGROUND: We designed a straightforward method for discriminating circulating Leishmania populations in the Indian subcontinent (ISC). Research on transmission dynamics of visceral leishmaniasis (VL, or Kala-azar) was recently identified as one of the key research priorities for elimination of the disease in the ISC. VL in Bangladesh, India, and Nepal is caused by genetically homogeneous populations of Leishmania donovani parasites, transmitted by female sandflies. Classical methods to study diversity of these protozoa in other regions of the world, such as microsatellite typing, have proven of little use in the area, as they are not able to discriminate most genotypes. Recently, whole genome sequencing (WGS) so far identified 10 different populations termed ISC001-ISC010.METHODOLOGY / PRINCIPLE FINDINGS: As an alternative to WGS for epidemiological or clinical studies, we designed assays based on PCR amplification followed by dideoxynucleotide sequencing for identification of the non-recombinant genotypes ISC001 up to ISC007. These assays were applied on 106 parasite isolates collected in Nepal between 2011 and 2014. Combined with data from WGS on strains collected in the period 2002-2011, we provide a proof-of-principle for the application of genotyping to study treatment outcome, and differential geographic distribution.CONCLUSIONS / SIGNIFICANCE: Our method can aid in epidemiological follow-up of visceral leishmaniasis in the Indian subcontinent, a necessity in the frame of the Kala-azar elimination initiative in the region.
AB - BACKGROUND: We designed a straightforward method for discriminating circulating Leishmania populations in the Indian subcontinent (ISC). Research on transmission dynamics of visceral leishmaniasis (VL, or Kala-azar) was recently identified as one of the key research priorities for elimination of the disease in the ISC. VL in Bangladesh, India, and Nepal is caused by genetically homogeneous populations of Leishmania donovani parasites, transmitted by female sandflies. Classical methods to study diversity of these protozoa in other regions of the world, such as microsatellite typing, have proven of little use in the area, as they are not able to discriminate most genotypes. Recently, whole genome sequencing (WGS) so far identified 10 different populations termed ISC001-ISC010.METHODOLOGY / PRINCIPLE FINDINGS: As an alternative to WGS for epidemiological or clinical studies, we designed assays based on PCR amplification followed by dideoxynucleotide sequencing for identification of the non-recombinant genotypes ISC001 up to ISC007. These assays were applied on 106 parasite isolates collected in Nepal between 2011 and 2014. Combined with data from WGS on strains collected in the period 2002-2011, we provide a proof-of-principle for the application of genotyping to study treatment outcome, and differential geographic distribution.CONCLUSIONS / SIGNIFICANCE: Our method can aid in epidemiological follow-up of visceral leishmaniasis in the Indian subcontinent, a necessity in the frame of the Kala-azar elimination initiative in the region.
KW - Journal Article
U2 - 10.1371/journal.pntd.0005420
DO - 10.1371/journal.pntd.0005420
M3 - A1: Web of Science-article
C2 - 28249021
VL - 11
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
SN - 1935-2727
IS - 3
M1 - e0005420
ER -