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. 2001 Jul;8(4):828-31.
doi: 10.1128/CDLI.8.4.828-831.2001.

Real-time PCR as a new tool for quantifying Leishmania infantum in liver in infected mice

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Real-time PCR as a new tool for quantifying Leishmania infantum in liver in infected mice

S Bretagne et al. Clin Diagn Lab Immunol. 2001 Jul.

Abstract

The parasitic loads of mouse livers experimentally infected with Leishmania infantum were determined using a double real-time quantitative PCR test targeted to the parasite DNA polymerase gene and to the mouse brain-derived neutrophic factor gene. The Leishmania DNA copy number was normalized to the number of mouse gene copies in order to quantify the former independently of liver weight. The correlation coefficient with the microtitration method was 0.66. This PCR assay can be considered for experimental pharmaceutical studies.

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Figures

FIG. 1
FIG. 1
Amplification plots and a standard curve obtained with the TaqMan Leishmania infantum PCR test. (A) Serial 10-fold dilution of L. infantum DNA from 1 to 106 ng per reaction (5 to 5 ×ばつ 106 copies/reaction); the amplification curves shift to the right as the input target quantity is reduced, since reactions with fewer target molecules require more amplification cycles to produce a detectable quantity of reporter molecules than do reactions with more target molecules. (B) Standard curve obtained by plotting the Ct against the input target quantity, with the latter plotted on a common log scale. Ct represents the fractional cycle number reflecting a positive PCR result differentiated from the background noise.
FIG. 2
FIG. 2
Correlation between log-transformed individual values of liver parasite burden determined by in vitro micotitration and quantitative PCR test.

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