Abstract
Visceral leishmaniasis, a vector-borne disease caused by Leishmania donovani and Leishmania infantum, currently affects 12 million individuals in 88 countries. In the present study, a real-time PCR (rt-PCR) assay has been optimized and validated against 2 other routine methods, i.e., microscopy and limiting dilution culture assay, to estimate parasite load in the liver of infected Syrian hamsters (Mesocricetus auratus). A set of specific primers amplified a 116-bp target template of the kinetoplastid DNA of L. donovani in a SYBR® Green-based rt-PCR assay. To assess the methods, we tested 2 anti-leishmanial compounds belonging to the class of arylimidamides, DB745 (2,5-bis[2-ethoxy-4-(2-pyridylimino)aminophenyl]furan) and DB766 (2,5-bis[2-(2-propoxy)-4-(2-pyridylimino)aminophenyl]furan) for efficacy in vivo in Syrian hamsters infected with L. donovani promastigotes. Parasite load was quantified in liver by all 3 methods and was found comparable. Of the 3 methods, rt-PCR was the fastest and most convenient, sensitive, and reproducible method.
| Original language | English |
|---|---|
| Pages (from-to) | 145-150 |
| Number of pages | 6 |
| Journal | Journal of Parasitology |
| Volume | 99 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Feb 2013 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Parasitology
- Ecology, Evolution, Behavior and Systematics
Cite this
Real-time PCR to quantify Leishmania donovani in hamsters. / Srivastava, Anuradha; Sweat, J. Mark; Azizan, Azliyati; Vesely, Brian; Kyle, Dennis E.
In: Journal of Parasitology, Vol. 99, No. 1, 02.2013, p. 145-150.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Real-time PCR to quantify Leishmania donovani in hamsters
AU - Srivastava, Anuradha
AU - Sweat, J. Mark
AU - Azizan, Azliyati
AU - Vesely, Brian
AU - Kyle, Dennis E.
PY - 2013/2
Y1 - 2013/2
N2 - Visceral leishmaniasis, a vector-borne disease caused by Leishmania donovani and Leishmania infantum, currently affects 12 million individuals in 88 countries. In the present study, a real-time PCR (rt-PCR) assay has been optimized and validated against 2 other routine methods, i.e., microscopy and limiting dilution culture assay, to estimate parasite load in the liver of infected Syrian hamsters (Mesocricetus auratus). A set of specific primers amplified a 116-bp target template of the kinetoplastid DNA of L. donovani in a SYBR® Green-based rt-PCR assay. To assess the methods, we tested 2 anti-leishmanial compounds belonging to the class of arylimidamides, DB745 (2,5-bis[2-ethoxy-4-(2-pyridylimino)aminophenyl]furan) and DB766 (2,5-bis[2-(2-propoxy)-4-(2-pyridylimino)aminophenyl]furan) for efficacy in vivo in Syrian hamsters infected with L. donovani promastigotes. Parasite load was quantified in liver by all 3 methods and was found comparable. Of the 3 methods, rt-PCR was the fastest and most convenient, sensitive, and reproducible method.
AB - Visceral leishmaniasis, a vector-borne disease caused by Leishmania donovani and Leishmania infantum, currently affects 12 million individuals in 88 countries. In the present study, a real-time PCR (rt-PCR) assay has been optimized and validated against 2 other routine methods, i.e., microscopy and limiting dilution culture assay, to estimate parasite load in the liver of infected Syrian hamsters (Mesocricetus auratus). A set of specific primers amplified a 116-bp target template of the kinetoplastid DNA of L. donovani in a SYBR® Green-based rt-PCR assay. To assess the methods, we tested 2 anti-leishmanial compounds belonging to the class of arylimidamides, DB745 (2,5-bis[2-ethoxy-4-(2-pyridylimino)aminophenyl]furan) and DB766 (2,5-bis[2-(2-propoxy)-4-(2-pyridylimino)aminophenyl]furan) for efficacy in vivo in Syrian hamsters infected with L. donovani promastigotes. Parasite load was quantified in liver by all 3 methods and was found comparable. Of the 3 methods, rt-PCR was the fastest and most convenient, sensitive, and reproducible method.
UR - http://www.scopus.com/inward/record.url?scp=84873658806&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84873658806&partnerID=8YFLogxK
U2 - 10.1645/GE-3221.1
DO - 10.1645/GE-3221.1
M3 - Article
VL - 99
SP - 145
EP - 150
JO - Journal of Parasitology
JF - Journal of Parasitology
SN - 0022-3395
IS - 1
ER -