Abstract
The production of gametocytes in field isolates of Plasmodium falciparum is not clearly understood even though the gametocytes are crucial for disease transmission. Samples collected during the malaria transmission period from two different regions of India were cultured in vitro for gametocyte production and analyzed by PCR and RT-PCR assay for Pfs25 gene. A total of 20 P. falciparum field isolates were collected which showed varying intensity of in vitro gametocyte production. The isolates which produced mature gametocytes in vitro also had an increase in their Pfs25 expression, indicating that the gametocyte produced is directly proportional to the Pfs25 gene expression. The expression ranged from 0.32 to 4.56 fold in field isolates when compared to the reference strain. The in vitro gametocyte production in fresh field isolates directly correlated with the expression of Pfs25 gene in these isolates as shown by ANOVA test.
Highlights
The malaria parasite lifecycle requires the production of gametocytes and transmission of gametocytes from human host to the vector
In two isolates (RNC60 and RAP4) no gametocyte production was seen inspite of their adaptation in vitro and the asexual stages identified in smears
It was observed that the culture adapted field isolates during gametocyte production in vitro showed an increased expression of Pfs25 gene when compared to a gametocyte producing reference strain (NF54)
Summary
The malaria parasite lifecycle requires the production of gametocytes and transmission of gametocytes from human host to the vector. This continuous spread of malaria causes 200-300 million clinical cases and 0.7 million deaths each year and P. falciparum is considered to be the deadliest of the human parasites [1]. The gametocytes are present at much lower concentrations compared to the asexual stages and may not be observed by microscopy [3]. Gametocytes ensure the transmission of malaria to the mosquito they are not responsible for clinical symptoms. The common anti-malarial drugs are not able to clear all gametocytes thereby mediating transmission to mosquitoes despite clearance of other erythrocytic stages [4]. With the advent of available newer technologies for detection of gametocytes and its sub patent population several molecular detection methods are used such as real-time quantitative nucleic acid sequence-based amplification (QT-NASBA), real-time polymerase chain reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP) [5,6,7,8]
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