Fig 1.
Workflow of the study.
Fig 2.
The one-dimensional (1D) ddPCR results from Bio-Rad QX100TM Droplet Reader of 18S rRNA Plasmodium genus detection and quantification assay.
The different amplitudes of positive droplets were observed when different PCR annealing temperatures were applied (A10: 65°C, D10: 62°C, E10: 60°C, F10: 59°C, H10: 57°C, F09: 60°C for negative control). Threshold for positive detection is 2500.
Fig 3.
The 95% probability of detecting parasitaemia as low as 10.674 parasites/mL when high volume of blood samples was applied with Plasmodium genus ddPCR assay.
Fig 4.
The linearity of ddPCR assays for 18S rRNA Plasmodium genus detection and quantification.
Linearity of ddPCR assay is shown using 5-fold serially diluted FACS concentrated DNA (from 2,000, 400, 80, 16 parasites/mL, respectively) and 18S rRNA concentration from ddPCR assay (copies/mL); R2 = 0.9986.
Fig 5.
The column bar graphs show 18S rRNA copies/mL of blood obtained from single reaction ddPCR and duplex ddPCR assays.
Mean 95% CI of 18S rRNA of P. falciparum (5a), P. vivax (5b), P. malariae (5c) and P. ovale (5d).
Table 1.
Identification of mixed infection by duplex ddPCR assay in comparison to real-time (performed in quadruplicate).
Table 2.
Diagnosis sensitivity and specificity of ddPCR assay for Plasmodium genus and species detection.