1.Detection of Plasmodium falciparum by using magnetic nanoparticles separa-tion-based quantitative real-time PCR assay
Fei WANG ; Yin TIAN ; Jing YANG ; Fujun SUN ; Ning SUN ; Biyong LIU ; Rui TIAN ; Guanglu GE ; Mingqiang ZOU ; Congliang DENG ; Yi LIU
Chinese Journal of Schistosomiasis Control 2014;(5):522-525,530
Objective To establish a magnetic nanoparticles separation-based quantitative real-time PCR(RT-PCR)assay for fast and accurate detection of Plasmodium falciparum and providing a technical support for improving the control and preven-tion of imported malaria. Methods According to the conserved sequences of the P. falciparum genome 18SrRNA,the species-specific primers and probe were designed and synthetized. The RT-PCR was established by constructing the plasmid standard , fitting the standard curve and using magnetic nanoparticles separation. The sensitivity and specificity of the assay were evaluat-ed. Results The relationship between the threshold cycle(Ct)and logarithm of initial templates copies was linear over a range of 2.5×101 to 2.5×108 copies/μl(R2=0.999). Among 13 subjects of entry frontier,a P. falciparum carrier with low load was de-tected by using the assay and none was detected with the conventional examinations(microscopic examinations and rapid tests). Conclusion This assay shows a high sensitivity in detection of P. falciparum,with rapid and accurate characteristics,and is especially useful in diagnosis of P. falciparum infectors with low parasitaemia at entry-exit frontier ports.
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