Carestart malaria

Bilirubin was measured using a FUJI DRI-CHEM 4000i automated system. Malaria was diagnosed by Rapid Diagnostic Test (RDT; CareStart™ Malaria, Access Bio, Somerset, NJ) and standard microscopy methodology. In a subgroup of newborns, malaria was assessed using ultrasensitive RDTs (Alere Malaria Ag P.f., Abbott, Chicago, IL). Hemoglobin and HCT were measured by HemoCue^® 301 (Angelholm, Sweden) and Hawksley Haematospin 1400 (Hawksley & Sons, Ltd., Lancing, United Kingdom), respectively. Anemia in the mother was defined as mild, moderate, or severe using WHO cutoffs.²² Polycythemia in the newborn was defined as venous HCT ≥ 65% or Hb ≥ 22 g/dL.²³ The ABO group and Rhesus type were assessed by the Beth-Vincent and Coombs tests, respectively. Glucose-6-phosphate dehydrogenase deficiency was assessed by RDT (CareStart™ G6PD Access Bio) and Fluorescent Spots Test (FST; R&D Diagnostic Ltd., Papagos, Greece). A dried blood spot was collected from the mother and her newborn(s) for identification of HbS (rs334) and G6PD 202G>A (rs1050828) from genomic DNA, using published DNA extraction²⁴ (link) and the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) methods.²⁵ (link)^,26 (link)

Madang is located on the north coast of Papua New Guinea (Fig. 1a). P. falciparum and P. vivax are highly endemic, and P. malaria and P. ovale are also present in the study area^{38 (link)}. Capillary blood samples were collected by qualified nurses or community health workers at Yagaum Rural Hospital located outside of Madang and at Madang Town Clinic in 2017–2019.
Blood samples originated from suspected malaria cases presenting as outpatients at the respective clinics. An RDT (CareStart^® Malaria, Accessbio Inc., USA) and two blood slides for LM were prepared at enrollment. All RDT positive patients received standard antimalarial treatment according to PNG malaria treatment guidelines. This study received ethical clearance by the PNG Institute of Medical Research (PNGIMR) Institutional Review Board and the PNG Medical Research Advisory Committee (MRAC, #16.45).
Samples from n = 12 long-term malaria-free and n = 20 malaria-naïve volunteers were also measured using RMOD and included in the analysis of the present study.
All tests were carried out independently and blinded to the results of each other. Written informed consent was provided by all study participants and, in case of minor patients, from their parents or guardians, in addition to assent from the minor patients.

Rapid Diagnostic Test (RDT) of malaria was performed on site using CareStart™ Malaria (Access Bio, Inc., New Jersey, USA) and parasite density was estimated microscopically from the positive samples as previously described [12 (link)].