Intermittent screening and treatment with artemether-lumefantrine versus intermittent preventive treatment with sulfadoxine-pyrimethamine for malaria in pregnancy: A facility-based, open-label, non-inferiority trial in Nigeria

Background: The spread of SP resistance may compromise the effectiveness of intermittent preventive treatment of malaria in pregnancy (MiP) with sulfadoxine-pyrimethamine (IPTp-SP) across Africa. However, there is no recommended alternative medicine for IPTp or alternative strategy for prevention of MiP. This poses problems for the prevention of MiP. This study investigated, whether screening with a rapid diagnostic test for malaria at routine antenatal clinic attendances and treatment of only those who are positive (intermittent screening and treatment) with artemether-lumefantrine is as effective and safe as IPTp-SP in pregnant women. Methods: During antenatal clinic sessions at the General Hospital Calabar, Nigeria, held between October 2013 and November 2014, 459 pregnant women were randomized into either the current standard IPTp-SP or intermittent screening and treatment with artemether-lumefantrine (ISTp-AL). All women received a long-lasting insecticide-treated net at enrolment. Study women had a maximum of four scheduled visits following enrolment. Haemoglobin concentration and peripheral parasitaemia were assessed in the third trimester (36-40 weeks of gestation). Birth weight was documented at delivery or within a week for babies delivered at home. Results: In the third trimester, the overall prevalence of severe anaemia (Hb < 8 g/dl) and moderate (8-10.9 g/dl) anaemia was 0.8 and 27.7%, respectively, and was similar in both treatment groups (p = 0.204). The risk of third-trimester severe anaemia did not differ significantly between both treatment arms (risk difference - 1.75% [95% CI - 4.16 to 0.66]) although the sample was underpowered for this outcome due to several participants being unavailable to give a blood sample. The risk of third-trimester maternal parasitaemia was significantly lower in the ISTp-AL arm (RD - 3.96% [95% CI - 7.76 to - 0.16]). The risk of low birthweight was significantly lower in the ISTp-AL arm after controlling for maternal age, gravidity and baseline parasitaemia (risk difference - 1.53% [95% CI - 1.54 to - 1.15]). Women in the ISTp-AL arm complained of fever more frequently compared to women in the IPTp-SP arm (p = 0.022). Conclusions: The trial results suggest that in an area of high malaria transmission with moderate sulfadoxine-pyrimethamine resistance, ISTp with artemether-lumefantrine may be an effective strategy for controlling malaria in pregnancy. Participation in the study was voluntary; subjects were free to withdraw from the study at any time, and this did not affect their access to or quality of care. Information obtained from all subjects was treated as confidential. The trial was conducted under the provisions of the Declaration of Helsinki in its most recent form (2013) and in accordance with Good Clinical Practices guidelines set up by the WHO and by the International Conference on Harmonization. The purpose of the study, the procedures to be followed and the potential risks, as well as benefits of participation, were explained to all participants. Information sheets and consent forms were provided to them for their review. Each participant signed an informed consent to participate in the research study. The study protocol and informed consent forms were reviewed and approved by the Cross River Health Research Ethics Committee, Calabar, Nigeria and the Ethics Board of the Medical Center of the University of Munich (LMU), Munich, Germany. The trial was prospectively registered with the Pan African Clinical Trials Registry (PACTR201308000543272). This study was an individually randomized, two-arm, observer-blinded, parallel-group, non-inferiority trial. The trial was undertaken between October 2013 and November 2014 to investigate whether screening for malaria with RDT and treating all women with a positive result using AL was not inferior to intermittent preventive treatment with SP in the prevention of anaemia in pregnancy. Eligible and consenting pregnant women were randomly assigned (1:1) to one of the two study arms (1) LLIN plus IPTp-SP or (2) LLIN plus ISTp-AL. The study was conducted in Calabar, Cross River State in South-East Nigeria. In the 2006 Population and Housing Census, Cross River state was made up of 1,471,967 males and 1,421,021 females with an annual growth rate of 2.9%. The projected population for 2015 is 3,783,085. The climate in Calabar is tropical-humid with wet and dry seasons, with average temperatures ranging between 15 and 30 °C and the annual rainfall between 1300 and 3000 mm. The vegetation in Calabar is mangrove swamp forest. Malaria transmission in this area is intense and perennial but with a peak in the rainy season, and Plasmodium falciparum is the predominant malaria-causing species [5, 14]. Previous studies have reported resistance to CQ and SP in Calabar to be over 80% [15]. Anopheles gambiae is the predominant vector species [16]. There is no information available on entomological inoculation rate (EIR) in the study area, but an EIR of about 259 infectious bites per person-year has been reported from Odukpani, a neighbouring area [17]. National HIV prevalence is reported to be about 3.4% with a higher prevalence of 5.5% in the region where the trial was conducted [18]. The trial was conducted at the antenatal clinics of the General Hospital in Calabar, Nigeria. The General Hospital is the largest government-owned secondary health facility in the city and caters to the health needs of the majority of the inhabitants. Since August 2009, pregnant women and children under 5 years of age receive free medical care as part of a funded welfare program by the Cross River State government. The average annual antenatal clinic attendance and births at the hospital are 16,550 and 3100 respectively. In the study area, the proportion of births attended by unskilled personnel has been estimated to be about 85% [19, 20]. In a health demographic and surveillance area 15 km from the trial site, only about 15% of births take place in health facilities [21]. The study population comprised pregnant women of all parities who presented at the antenatal clinics at their first booking. Antenatal care clinics were conducted 3 days per week; Wednesdays for first visits, then Mondays and Thursdays for follow-up visits. Women were screened for eligibility and invited to participate in the study if they met the criteria. Eligible participants were HIV-negative pregnant women between 16 and 24 weeks’ gestation at their first booking, no history of receiving intermittent preventive treatment with sulfadoxine–pyrimethamine during the pregnancy, resident in the study area, and willing to have a supervised delivery. Women with any illness requiring hospital admission (including severe malaria as defined by WHO), high-risk pregnancies, severe anaemia (Hb < 6 g/dl), known G6PD deficiency, a history of sensitivity to SP, lumefantrine or an artemisinin, or those unwilling to give consent were excluded. The sample size was calculated based on the assumption that the prevalence of severe anaemia in the third trimester of pregnancy in the IPTp-SP arm of the study would be at least 3% based on findings from a previous study undertaken in West Africa [13]. Due to widespread resistance to SP, it was hypothesized that the prevalence of severe anaemia would be significantly lower in the LLIN plus ISTp-AL arm compared to the LLIN plus IPTp-SP arm. To establish that ISTp-AL was not inferior to IPTp-SP, it was necessary to show that the differences in the proportion of women with severe anaemia between both arms would not be more than 5%, a difference which would be of clinical and public health importance. To meet these criteria with 80% statistical power and allowing for 20% loss to follow-up it was calculated that 230 women were needed in each study arm giving a total sample size of 460. All pregnant women presenting for their first antenatal care visit at the General Hospital were invited to attend group information sessions and were screened for their eligibility. After women had provided written informed consent, they were randomised to one of the two treatment groups (ISTp-AL or IPTp-SP arm) in computer-generated permuted blocks of ten. At enrolment, an eligible pregnant woman (16–24 weeks gestational age) was asked by a designated study nurse to pick a slip from the opaque envelope. The slip contained the treatment group the woman had been assigned to. The study arm women belonged to was not identifiable by the identification numbers given to them. Also, the principal investigator and outcome assessors (midwives and microscopist) were blinded to the randomization process and treatment allocation to prevent bias in outcome assessment. Women who declined to participate in the trial were treated with the routine standard of care, IPTp-SP according to the national guidelines. Enrolled study women were randomized to either the ISTp-AL or IPTp-SP arm. All study women received a LLIN, which they were encouraged to use throughout the pregnancy. Also, they received a daily supplement of folic acid (4 mg) and ferrous sulphate (200 mg) tablets according to national guidelines. HIV screening was offered to all study women as part of the routine antenatal services recommended in Nigeria with an option for treatment, but the results of HIV screening were not available to the study team at the time of enrolment. At enrolment, a finger prick blood sample was obtained for determination of haemoglobin concentration, preparation of blood smears (thin and thick films) for malaria parasite counts and preparation of dried blood spots (DBS) on filter paper. Women in the IPTp-SP arm received an initial dose of SP (1500 mg sulfadoxine/75 mg pyrimethamine) as a single dose. Pregnant women in ISTp-AL arm were screened for malaria infection with an SD Bioline rapid diagnostic test (RDT), a histidine-rich protein-2 (HRP-II) antigen and Plasmodium lactate dehydrogenase (pLDH) (Pan) antigen RDT kit. The RDTs were purchased from Codex Pharma Limited, Nigeria, as needed and stored following the manufacturer’s instructions. The tests were also performed and interpreted by the study team following the manufacturer’s instructions. Women in the ISTp-AL arm were treated with artemether–lumefantrine (20 mg artemether/120 mg lumefantrine) given as a 6-dose course, administered twice daily for 3 days if the RDT was positive. Study women were advised on the time and mode of administration for the 3 days treatment taken at home unobserved. Women in the ISTp-AL arm received no anti-malarial treatment if their RDT results were negative. Laridox® (sulfadoxine–pyrimethamine, IPCA Laboratories Ltd, India) and Coartem® (artemether–lumefantrine, Novartis Pharma, Switzerland) were used for this trial. Study women were asked to return for follow-up antenatal care visits and IPTp-SP or screening with RDT at 24, 32 and 36 weeks of gestation if they were in the ISTp-AL group. At the next two follow-up visits (at 24 and 32 weeks of gestation), women in the IPTp-SP arm received SP while women in the ISTp-AL arm were screened with the RDT and, if positive, treated with AL. All women were followed-up by a study nurse through phone calls and at subsequent visits to document any complaints or adverse events. Study women were also provided with a mobile phone number, which they could call to arrange unscheduled visits. Blood samples were obtained from all women in the third semester (36–40 weeks of gestation) before delivery for the determination of haemoglobin concentration and the preparation of blood smears. However, the smears were read retrospectively, and so the results were not available at the point of care. At delivery, placental smears were collected on microscopy slides to determine the prevalence of placental malaria. Study woman who came to the hospital with a history of fever or other symptoms of malaria between scheduled antenatal care visits was screened for malaria with RDT and if positive for malaria, treated with quinine (30 mg/kg daily for 5 days) regardless of treatment group according to national policy. Birth weight was measured by a midwife who was unaware of the treatment group of the woman whom she was attending. The occurrence of miscarriages, stillbirths, neonatal deaths and the presence of congenital abnormalities were also recorded by midwives. Women were invited to re-attend at 6 weeks postpartum with their baby. A blood film was obtained from the mother. Her haemoglobin concentration was also measured at this time. Also, any neonatal adverse events were documented. A team of nurses, lab scientists was dedicated to the follow-up of women who did not present for delivery at the General hospital within 1 month of the estimated delivery date to establish pregnancy outcome. Labelled blood and impression smears were air-dried and stained with 4% Giemsa for 30 min. Thick smears were used to count the number of asexual parasites per 200 leukocytes, assuming 8000 leukocytes/μl of blood; slides were declared negative if no parasite was seen after examination of 100 high power fields. All blood films were read, and parasitaemia was quantified by two experienced and blinded microscopists independently. If there was a discrepancy in the findings in a slide between the two microscopists (positive or negative or a ≥ 50% difference in parasite density) a third, more senior microscopist read the slide and their reading was deemed to be the correct reading. Haemoglobin (Hb) concentration was measured using Hb 301 Hemocue (HemoCue AB, Angelholm, Sweden; accuracy of 0.1 g/dl). Case report forms were completed for maternal, and neonatal adverse events (AEs) detected at scheduled antenatal and postpartum visits, at unscheduled visits and delivery. The clinician on-call was alerted by the nursing team upon detection of a possible serious adverse event (SAE). AEs were considered SAEs if they fulfilled one of the following criteria: the event resulted in death, led to hospitalization, was a congenital abnormality, was life-threatening, caused disability, or was deemed serious for other medical reasons. Serious adverse event reports were completed for study women who experienced miscarriages or stillbirths. Significant reductions in haemoglobin concentration (≤ 6 g/dl) after enrolment were also considered SAEs. Reports of drug side effects at each treatment course were considered as separate AEs. Stata version 12 (StataCorp, College Station, Texas) was used for data analyses. The primary objective of the trial was to show that the risk of third-trimester severe anaemia (Hb ≤ 8 g/dl) in the ISTp-AL group was no more than 5% greater than in the IPTp-SP group. Secondary objectives were to demonstrate that the risks of low birth weight (< 2500 g), intrauterine deaths/stillbirths, spontaneous abortions, neonatal and maternal mortality were not significantly higher in women in the ISTp-AL group than in women who received IPTp-SP. The principal analysis of both primary and secondary outcomes was according-to-protocol (ATP), but a modified intention-to-treat (mITT) analysis was also undertaken. In the ATP analysis, only data from women who remained within their randomization group and had a record of the primary outcome were included. This implied that study women had to have received two courses of SP (IPTp-SP arm) or been screened twice using an RDT at scheduled visits (ISTp-AL arm) and also, had a measurement for the primary outcome (haemoglobin level at 36–40 weeks gestation). In the modified intent-to-treat analysis (mITT), data from women who received an initial treatment of IPTp or had an initial screening test done and had a record for the primary outcome or the outcome of interest recorded was included. This implied that study women had to have received one course of SP (IPTp-SP arm) or been screened once using an RDT at scheduled visits (ISTp-AL arm) and also, have a measurement for the primary outcome or outcome of interest. The proportion of ATP and mITT populations experiencing each outcome for the trial arms, and the associated 2-sided 95% CI for the risk difference, was estimated using the generalized linear model. To declare non-inferiority with a significance level of 0.05, the upper boundary of the 2-sided 95% CI for the estimated treatment effect had to be below the pre-defined non-inferiority margin (∆) of 5%. Gestational age at enrolment, being primigravidae, baseline anaemia and parasitaemia were controlled for using binomial regression. Only birth weights from singleton pregnancies of live births ≥ 22 weeks’ gestation with no congenital abnormality and measured within 6 days of delivery were included in birth weight analyses. p-values less than 0.05 were considered statistically significant.