Table 1.
Primer sequences of nested PCR.
Table 2.
Primer & probe sequence of real time PCR.
Table 3.
Demographic parameters of study participants.
Fig 1.
Technical performance and range of detection of the Leishmania real-time PCR assay.
In (A) DNA was extracted from serial dilutions of cultured L. donovani, ranging from 1 x 105 to 1x 10−1/ ml, and subjected to real time PCR. Amplification curves are shown for each sample, with each parasite concentration depicted by a differing color. In (B) the mean Ct values are plotted from triplicates tested against serial dilutions containing 10ng to 10fg of L.donovani genomic DNA per reaction. Each point represents the Ct of an individual sample, with the plot of Ct values and parasite equivalent fitting a linear function (R2 _0.998).
Table 4.
Repeatability and reproducibility of the real time PCR assay.
Fig 2.
Parasite DNA can be detected in active, but not cured, disease states.
In (A), parasite burden in blood samples from VL (n = 40), VL patients deemed to be cured by symptomatic response to treatment (n = 40) and RVL patients (n = 10) were measured. Each point indicates the data obtained from each individual sample. In (B), parasite burden in skin samples from PKDL patients before or after treatment (n = 40) were measured. Each point indicates the data obtained from each individual sample, and before and after data for each patient are linked by the connecting line.
Table 5.
Sensitivity and specificity of Ln-PCR and real time PCR in detecting L. donovani in clinical samples.
Fig 3.
Receiver operating characteristic (ROC) curve for Ln-PCR for the detection of Leishmania parasites in clinical samples.