Fig. 4

Quantitative correlation between gene copy number and threshold cycle of the allele specific real-time PCR assay. A, B HGPRT plasmid was serially diluted from 10² to 10⁸ copies/reaction and subjected to allele specific qPCR. Linear regression of Ct vs lg copy number of HGPRT plasmid were generated using single and multi-species templates for L. donovani (A) and L. infantum (B) detection, individually. C SPDSYN plasmid was serially diluted from 10² to 10⁸ copies/reaction and subjected to allele specific qPCR. Linear regression of Ct vs lg copy number of SPDSYN plasmid were generated using single and multi-species templates for L. major detection. D, E SPDSYN plasmid was serially diluted from 10³ to 10⁹ copies/reaction and subjected to allele specific qPCR. Linear regression of Ct vs lg copy number of SPDSYN plasmid were generated using single and multi-species templates for L. tropica (D) and L. donovani/infantum (E) detection, respectively. ΔRn = Rn (normalized reporter)-baseline. Ct Cycle threshold