A dried blood sample on filter paper is suitable for detecting Plasmodium falciparum gametocytes by reverse transcription polymerase chain reaction

In vivo and in vitro gametocyte production of Plasmodium falciparum isolates from Northern Thailand

“…We refer to the five allelic types of pfg377 as A, AB, B, C and D, as distinguished by size polymorphism of PCR amplified fragments. Alleles A, B, C and D were the same size as those previously described from Vietnamese malaria patients in 2002 (Maeno et al, 2008), whereas allele AB shared a similar size to an allele previously described from Africa, for which no nucleotide sequence was previously determined. The sizes of RT-PCR products of alleles A, AB, B, C and D, as visualized by agarose gel electrophoresis, were 411, 390, 369, 348 and 306 bp, respectively.…”

“…The recent application of more sensitive molecular detection methods such as real-time nucleic acid sequence based amplification (QT-NASBA) (Schneider et al, 2005), reverse transcription-PCR (RT-PCR) (Maeno et al, 2008) and reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) (Buates et al, 2010) have revealed that the proportion of parasite-infected individuals who harbour gametocytes is much higher than previously thought, and has led to a much clearer picture of the patterns of gametocyte carriage amongst infected individuals (Ouedraogo et al, 2009). The limit of detection of gametocytes by thicksmear microscopy is in the range of five gametocytes per µl, while molecular techniques such QT-NASBA can detect as few as 0.02 gametocytes per µl (Schneider et al, 2007).…”

High specificity of semi-nested multiplex PCR using dried blood spots on DNA Banking Card in comparison with frozen liquid blood for detection of Plasmodium falciparum and Plasmodium vivax

“…Results obtained from a molecular epidemiologic study based on nested-PCR technique indicated that airdried spots onto Whatman chromatography filter paper resulted in a better outcome for detection of P. falciparum than thick Geimsa-stained blood films in microscopical examination (19). Maeno et al in a molecular-based P. falciparum gametocyte detection study suggested that blood sample spotted on the filter paper is a useful sampling for some fields of malaria studies (20). In another study, some authors reported that mRNA is a stable target for amplification by RT-PCR technique using 3-month stored dried blood at ambient temperature (18).…”

Assessing the Gametocyte Production in Field Isolates of Plasmodium falciparum

“…The culture was maintained continuously and Giemsa stained thin smears were examined at 1000X magnification under the microscope every 24 hours to monitor the parasitaemia levels and the asexual stages. The isolates were sub-cultured when the parasitaemia reached 4.0% and since it is known that 1 µl of blood contains 5x10 6 RBCs the percent parasitaemia was thus calculated [14].…”

“…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 now 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].…”