METHODS: Two hybridoma clones (Aa3c10, b10c1) were used for the production of ascites in BALB/c mice. Purification of monoclonal antibodies from the ascites was carried out using affinity columns. The thermal stability study of monoclonal antibodies was done by storing it at 37°C and 45°C for thirty days. The stored antibodies were analyzed using SDS-PAGE and flow-through device where the antigenantibody interaction was visualized by Protein A colloidal gold solution. Sensitivity was determined by endpoint dilution ELISA and the dissociation constant by competitive ELISA. Sensitive pair optimization was done by sandwich ELISA using biotinylated antibodies. Prototype preparation for lateral flow assay had a colloidal gold-based detection system.
RESULTS: Thermal stability experiments showed that both mAbs (Aa3c10; b10c1) are stable up to thirty days at 45°C while the commercially available mAbs were stable up to fifteen days only. Compared to commercial antibodies, the mAb Aa3c10, showed the highest sensitivity in end-point titre. In sensitive pair optimization, it was observed that the mAb, b10c1, as a detector and the mAb, Aa3c10, as a capture antibody showed the highest absorbance to detect 50pg/ml PfHRP2 antigen. The prototype formulation of lateral flow assay using the mAbs (Aa3c10; b10c1) showed good reactivity with WHO panel and no false-positive results were observed with twenty clinically negative samples and five P. vivax positive samples.
CONCLUSIONS: The novel monoclonal antibodies (Aa3c10, b10c1) against truncated PfHRP2, could be a strong potential candidates that can be included in making RDTs with better sensitivity and stability.
Method: The study involved secondary data analysis from the Malaysian Health and Adolescents Longitudinal Research Team (MyHeART) study. A closed cohort secondary data analysis was performed from the dynamic cohort of 528 adolescents (male = 151; female = 377) aged 13 years attending secondary school who were followed up at 15 and 17 years. Anaemia status was determined by haemoglobin level < 12g/dL based on FBC, and iron deficiency anaemia (IDA) was determined when the Mentzer Index < 13. A generalised estimating equation (GEE) was constructed to investigate the longitudinal relationship between nutritional status and lifestyle on anaemia status over five years.
Results: The trend of anaemia prevalence increased significantly across the age group (7•9%; 95% CI: 2•3-11•1, 13•9%; 95% CI: 10•8-15•7 and 15•8%; 95% CI: 3•8-23•1) at 13, 15 and 17 years, respectively, especially among females. The trend of anaemia prevalence among females, also increased significantly across the age group (11.1%;95% CI:6.7-17.8, 15.7%;95% CI:11.4-21.3, 23.1%;95% CI:16.8-31.0). A similar trend was noted for the prevalence of IDA among those who were anaemic (66•5%; 95% CI: 40•4-85•3, 72•2%;95% CI: 54•8-85•4, 76•3%; 95% CI: 59•2-87•7). A longitudinal analysis using GEE revealed that adolescents who did not meet the Recommended Nutrient Intake (RNI) for total iron intake per day were significantly associated with anaemia (RR=1•517;95% CI: 1•012-2•275; p=0•044) and IDA (RR=1•776;95% CI: 1•225-2•57; p= 0•002).
Conclusion: The overall trend of anaemia among adolescents is in increasing trend and anaemia is prevalent among female adolescents in this study. It is crucial to understand that the current fortification strategy may need to be revisited, and robust intervention programmes are necessary and should be sex specific.