Carestart

The study was conducted between 2013 and 2014 to assess the performance of BinaxNow (Binax Inc, Inverness Medical, ME, USA), SD Bioline (Standard Diagnostics Inc, Korea), Carestart (Access Bio Inc, Monmouth, USA), and First Response (Premier Medical Co Ltd, India). (1) BinaxNow, which targets both P. falciparum-specific HRP2 and a pan-malarial antigen, Plasmodium aldolase; (2) SD Bioline 05FK50, which targets P. falciparum-specific HRP2 only and 05FK60, which targets both P. falciparum-specific HRP2 and the pan-malaria antigen pLDH; (3) Carestart IFU G0131, which targets both P. falciparum-specific HRP2 and the pan malarial antigen, pLDH, IFU G0141, which targets P. falciparum-specific HRP2 only and IFU G0181, which targets both P. falciparum-specific HRP2 and pLDH; and, (4) First Response I13FRC, which targets P. falciparum-specific HRP2 and First Response I16FRC, which targets both P. falciparum-specific HRP2 and the pan-malarial antigen pLDH.
Participants were recruited to the study through the Kombewa HDSS. All patients, irrespective of their age and gender, presenting at the health facilities with a clinical suspicion of malaria were included in the study after taking written informed consent. The study design was in compliance with the STARD methodological guidelines for presentation of diagnostic studies [13 (link)].

Socio-demographics data, medical and obstetric history, the use of medication and insecticide-treated bed nets were collected using an individual case record format. Gestational age was determined by the last menstrual period (LMP) and the fundal height method, which is the standard of care in Tanzania. Weight was measured by digital weighing scale in kilograms (nearest 0.1 kg). Height was measured by a potable wooden scale in centimeter (nearest 0.1 cm). Temperature was recorded from the maternal armpit using a digital thermometer and fever was defined as a temperature of ≥37.5 °C. All recruited pregnant women were tested for HIV sero-status as a part of a routine ANC procedure. Peripheral finger-prick blood was collected for malaria detection and hemoglobin concentration determination. Asymptomatic malaria was detected by RDT (Care start, ACCESS BIO Somerset, NJ, USA) and real-time PCR (Applied Biosystems). Hemoglobin concentration was determined by HemoCue Hemoglobin 201+ analyzer (HemoCue AB Angelholm, Sweden).

The study was performed in the health district of Nanoro, central-west of Burkina Faso. Samples and data were collected in three peripheral health facilities (Nanoro, Nazoanga and Seguedin), and in the referral hospital, the Centre Médical avec Antenne Chirurgicale (CMA) Saint Camille of Nanoro. The only diagnostic tool available for the routine diagnosis of fever in patients seeking heath care in these clinics is a malaria RDT. The RDTs used in government clinics are one based on the detection of PfHRP2 (manufacturers: SD Bioline: Standard Diagnostics, Hagal-Dong, Korea; CareStart: Access Bio, Inc.). The management of uncomplicated medical cases, including uncomplicated malaria (positive malaria RDT), is done at the health facilities by nurses according to the national guideline of integrated management of childhood illness [14 ]. Suspected malaria infection is the first reason for consultation in children under-5 years of age in this region and predominately occurs during the rainy season between July and November. The annual prevalence of malaria in children under-5 years of age has previously been determined to be 49.7% [15 (link)].

All resident children were eligible for randomization when aged 6–59 months and with a height-for-age z-score in the range −3 SD to 1.5 SD. Children with haemoglobin concentration <70 g/L, signs of chronic illness, and those unlikely to remain permanently resident or comply with the supplementation for the duration of the trial, or whose parents or guardians declined consent, were excluded from the study. Venous blood samples were collected in EDTA tubes and centrifuged immediately. An aliquot of 90 μL erythrocyte sediment with the buffy coat was mixed with 90 μL phosphate-buffered saline and 180 μL of DNA stabilizing buffer (AS1; Qiagen, Hilden, Germany) and stored at 4°C for subsequent genotyping. Plasma samples were stored in liquid nitrogen in the field and at −80°C during transport and subsequent storage until biochemical analysis in The Netherlands. Haemoglobin concentration was measured in an aliquot of whole blood by a haematology analyser (Sysmex KX21, Kobe, Japan). Plasmodium infection was detected in fresh blood by rapid dipstick test (CareStart, Access Bio, Monmouth Jct, USA). Children with a positive test result were treated immediately with artemether-lumefantrine. The location of the child’s homestead was determined using a global positioning system. Further details about recruitment procedures are reported elsewhere [21 (link)].

Trial patients were hospitalized on days 1 to 3, and the trial visits took place on days 5, 8, 11, and 15 during the treatment period and on days 22, 29, 60, 90, 120, 150, and 180 during the follow-up period. Safety, clinical, laboratory, and parasitologic assessments were consistent with those in the phase 3 DETECTIVE trial.^{5 (link)}P. vivax genotyping and cytochrome P450 2D6 (CYP2D6) genotyping were performed as described in the phase 3 DETECTIVE trial.^{5 (link),14 -18 (link)} The incidence of genetically homologous or heterologous P. vivax infection was assessed with the use of previously published markers.^{15 (link)} G6PD phenotype was determined by means of quantitative spectrophoto-metric analysis (Trinity Biotech).^{5 (link)} In addition, a qualitative G6PD test (CareStart, AccessBio) was performed at screening, but the findings were not used to determine eligibility. G6PD genotype was determined for all female patients and for male patients who had a protocol-defined decrease in the hemoglobin level during the treatment period.^{19 (link),20 (link)}

Malaria infection was measured by rapid dipstick tests based on the detection of histidine-rich protein-2 and Plasmodium-specific lactate dehydrogenase (CareStart, product code G0121; Access Bio). In addition, Plasmodium falciparum-specific DNA was extracted from erythrocytes, and malaria parasite counts were detected at the Amsterdam Medical Centre, the Netherlands, based on a published protocol (29 (link)).