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Background  Compared to expert malaria microscopy, malaria biomarkers such as  Plasmodium falciparum  histidine rich protein-2 ( Pf HRP-2), and PCR provide superior analytical sensitivity and specificity for quantifying malaria parasites infections. This study reports on parasite prevalence, sick visits parasite density and species composition by different diagnostic methods during a phase-I malaria vaccine trial.    Methods  Blood samples for microscopy,  Pf HRP-2 and  Plasmodium  lactate dehydrogenase ( p LDH) ELISAs and real time quantitative PCR (qPCR) were collected during scheduled (n = 298) or sick visits (n = 38) from 30 adults participating in a 112-day vaccine trial. The four methods were used to assess parasite prevalence, as well as parasite density over a 42-day period for patients with clinical episodes.    Results  During scheduled visits, qPCR (39.9%, N = 119) and  Pf HRP-2 ELISA (36.9%, N = 110) detected higher parasite prevalence than  p LDH ELISA (16.8%, N = 50) and all methods were more sensitive than microscopy (13.4%, N = 40). All microscopically detected infections contained  P. falciparum , as mono-infections (95%) or with  P. malariae  (5%). By qPCR, 102/119 infections were speciated.  P. falciparum  predominated either as monoinfections (71.6%), with  P. malariae  (8.8%),  P. ovale  (4.9%) or both (3.9%).  P. malariae  (6.9%) and  P. ovale  (1.0%) also occurred as co-infections (2.9%). As expected, higher prevalences were detected during sick visits, with prevalences of 65.8% (qPCR), 60.5% ( Pf HRP -2  ELISA), 21.1% ( p LDH ELISA) and 31.6% (microscopy).  Pf HRP-2 showed biomass build-up that climaxed (1813±3410 ng/mL SD) at clinical episodes.    Conclusion    Pf HRP-2 ELISA and qPCR may be needed for accurately quantifying the malaria parasite burden. In addition, qPCR improves parasite speciation, whilst  Pf HRP-2 ELISA is a potential predictor for clinical disease caused by  P. falciparum .    Trial Registration  ClinicalTrials.gov  NCT00666380

plos one

Comparison of PfHRP-2/pLDH ELISA, qPCR and Microscopy for the Detection of Plasmodium Events and Prediction of Sick Visits during a Malaria Vaccine Study