Fig. 2

qPCR analysis of A. tetraptera DNA using LNA based-primers and LNA-TaqMan probe. a Schematic of the qPCR method using LNA-based primers and LNA-based TaqMan probe is shown. qPCR was conducted with the combination of the LNA-based primers and LNA-based TaqMan probe, which was modified with FAM, a fluorescent reporter at the 5′-end— and IBFQ, a fluorescence quencher at the 3′-end. The fluorescence of FAM within the probe disappeared by IBFQ during hybridization. The probe was cut using Taq polymerase during the extension reaction, which resulted in the emission of the reporter’s fluorescence. b qPCR was conducted using 100 copies of synthesized A. tetraptera DNA as template DNA in the presence of the indicated fluorescence reagents and primers. The detected RFU values were plotted and the representative data obtained from three independent experiments are shown. c qPCR was conducted using 3-fold serial dilutions of synthesized A. tetraptera DNA from 3000 to 1.37 copies as template DNA in the presence of the indicated fluorescence reagents and primers. The data of the copy numbers were converted to logarithm. The correlation coefficient indicates the relationship between the Ct value and the copy number of the template DNA. The data represent the mean ± SE obtained from 3 independent experiments. d qPCR was conducted using genomic DNA extracted from the faeces of A. tetraptera-infected mice in the presence of LNA-based TaqMan probes and LNA-based primers. The genomic DNA were used with a 3-fold serial dilution that ranges from 15,000 pg to 20.6 pg. The data of the copy numbers were converted to logarithm. The correlation coefficient indicates the relationship between the detected copy number of A. tetraptera DNA and the input genomic DNA. The dashed lines show the 95% confidence interval. The data represent the mean ± SE obtained from three independent experiments