Age and seasonal patterns of molecular force of blood-stage infection (_molFOB, Panel A) and incidence of clinical <i>P.</i><i>vivax</i> malaria (Panel B), and effects of ITN use on these parameters.

<p>Smooth splines from generalized additive models with single predictors and 95% confidence intervals.</p

Association of incidence of <i>P.</i><i>vivax</i> episodes with age in children with low (_molFOB: 0–10.7), medium (_molFOB: 10.7–18.0) and high average exposure (_molFOB: 18.1–39.0) to <i>P. vivax</i> infections.

<p>Incidence rate ratio and 95% confidence intervals for changes associated with 1 year increase in age.</p

Seasonality of <i>P.</i><i>vivax</i> clinical episodes (as compared to the first week of January, where incidence peaked) excluding _molFOB (blue) and adjusted for _molFOB (red).

<p>_molFOB accounts for approximately 50% of the seasonal variation in incidence. IRR = incidence rate ratio.</p

Association between total and IgG subclasses to 6 PvRBPs and protection against clinical malaria (density > 500/μL) in 224 young Papua New Guinean children.

<p>Data are plotted as exposure (molFOB), age, season and village of residency adjusted incidence rate ratios and 95% confidence intervals. Incidence rate ratios, 95% confidence intervals and P-values from GEE models. P < 0.05 were deemed significant. ****P < 0.001; ***P = 0.001; ** P > 0.001 to 0.01; *P > 0.01 to 0.05.</p

Effect of repeated sampling on molecular detection of parasite clones and on multiplicity of infection.

1<p>The highest value for MOI of either marker <i>msp1</i>F3 or MS16 was used.</p>2<p>Detectability refers to individual genotypes at day 1 versus day 2.</p

Erythrocyte-binding preferences of 6 PvRBPs.

<p>A: Bar charts showing the percentage of binding of CD71-PECy5 (control), PvRBP1a, PvRBP1b, PvRBP2a, PvRBP2b, PvRBP2cNB, PvRBP2-P2 to mature erythrocytes (not stained with thiazole orange, TO-) vs reticulocytes (stained with thiazole orange, TO+) populations. Error bars represent SEM of 7 or 9 independent repeats. B: Dot plots show the binding of six PvRBP proteins to an enriched reticulocyte population. Binding was detected using an anti-PvRBP rabbit IgG antibody followed by a secondary anti-rabbit Alexa 647 antibody.</p

Detection of PvRBPs by rabbit anti-PvRBP polyclonal antibodies by ELISA.

<p>Microtiter wells were coated with each PvRBP per well as shown by symbols on the left. Solid lines show specific anti-PvRBPs polyclonal antibodies (top label) added to each plate in a dilution series. The optical density (OD) was measured at 405 nm. Mean OD values from duplicated wells and standard deviation are shown.</p

Examples of results obtained on two consecutive days by PCR-capillary electrophoresis.

<p>Capillary electrophoresis chromatograms obtained with the <i>P. vivax</i> marker <i>msp1</i>F3 from two patients on two consecutive days. X-axis: size of PCR product in base pairs. Y-axis: relative fluorescent units. In patient 1 one clone was detected on day 1, and two additional clones were detected on day 2, combined MOI = 3. Patient 2 was negative on day 1, but one clone was found on day 2, combined MOI = 1.</p

Effect of repeated sampling on <i>P. falciparum</i> and <i>P. vivax</i> prevalence by light microscopy or PCR.

1<p>A sample was defined positive for <i>P. vivax</i> if any of the two markers <i>msp1</i>F3 or MS16 was amplified.</p>2<p>These samples are positive on day 1 for at least one marker and negative on day 2 for both markers.</p>3<p>These samples are negative on day 1 for both markers and positive on day 2 for at least one marker.</p>4<p>Detectability refers to PCR positivity at day 1 versus day 2.</p

Dynamics of parasite clones over 24 hours.

<p>Schematic overview of possible outcomes of 24 h bleeds. A sample is positive as soon as a parasite is detected on either day. Different colors of PCR results indicate different clones detected. The combined multiplicity of infection includes all clones detected in two corresponding bleeds.</p