Plasmodium that causes malaria disese is transmitted by the bite of an infected female Anopheles mosquito. Malaria causes high morbidity and mortality rates in sub-Saharan Africa compared to other parts of the world. Critical to malaria prevantion is the breaking of malaria transmission which is currently achieved by vector control methods or by chemotherapeutical approaches utilising antimalarial drugs such as primaquine. However, in many malaria endemic countries in Africa, including Zambia, primaquine use cannot be used due to less information on the prevalence of Glucose 6 phosphate dehydrogenase deficiency (G6PD), which causes haemolysis. Literature on the prevalence of (G6PD) in Sub-Saharan Africa, especially Zambia, is little. In malaria prevention, the importance of asymptomatic and subclinical parasitaemia which normally go along with gametocytaemia are not well stated in Zambia. The objectives of this study were to determine the prevalence of subclinical gametocytaemia and G6PD deficiency in high malaria transmission zones of Zambia as well as to assess the relationship between age and Plasmodium falciparum sexual stage Pfs25 gene in the study population. A total of 320 participants were enrolled into a cross sectional study from 5^th September 202022 to 29^th September 2023. Ethical clearance to conduct the study was obtained from Kesmonds International University Research Ethics Committee and informed consent process was completed with participants and or caregivers prior to enrolments. Finger-prick blood specimens were collected from all the 320 participants for the CareStart G6PD rapid diagnostic test (RDT) strip with assay buffer detecting conversion of pentose phosphate pathway co-enzyme nicotinamide adenine dinucleotide phosphate (NADPH) to its reduced form to detect G6PD enzyme deficiency, heat-fixed thick and alcohol-fixed thin films both stained in 10% Giemsa stain and examined microscopically to detect sexual and asexual forms of Plasmodium and identify Plasmodium species and Reverse Transcription-Polymerase Chain Reaction (PCR), conventional and nested PCR to detect Plasmodium falciparum sexual stage Pfs25 gene. Gametocytaemia prevalence in the high transmission zone was 2.8% (9/320) by microscopy and 9.1% (29/320) by PCR. Prevalence of G6PDd in high transmission zone was 2.5% (8/320), with seven (87.5%) of the G6PDd individuals, being males and only one (12.5%) was a female. The study reviewed 100% Plasmodium falciparum species. Presence of Plasmodium falciparum sexual stage Pfs25 gene did not differ significantly by age group. We demonstrate the presence of Plasmodium falciparum sexual stage Pfs25 gene in high malaria transmission zones indicating that malaria transmission still occurs in although the prevalence is expectedly higher in the high transmission zone. With the non-significant occurrence of Plasmodium falciparum sexual stage Pfs25 gene by age group, we conclude that all age groups are capable of transmission and attention must be paid to all age groups in the control and prevention programmes. We suggest that Primaquine use can be used in the high malaria transmission zones when G6PDd was absent. Though preliminary and notwithstanding the study limitations, the generated results on subclinical gametocytaemia and G6PDd profiles in high malaria transmission zones are important and will inform national policy in the ongoing current national malaria pevention activities. Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), Plasmodium falciparum, gametocytaemia (Pfs25) gene, glucose-6-phosphate dehydrogenase deficiency (G6PDd), high malaria transmission zones, malaria prevention, Zambia.