Detection of Plasmodium falciparum by using magnetic nanoparticles separa-tion-based quantitative real-time PCR assay
- VernacularTitle:纳米磁分离法结合实时荧光定量PCR检测恶性疟原虫的研究
- Author:
Fei WANG
;
Yin TIAN
;
Jing YANG
;
Fujun SUN
;
Ning SUN
;
Biyong LIU
;
Rui TIAN
;
Guanglu GE
;
Mingqiang ZOU
;
Congliang DENG
;
Yi LIU
- Publication Type:Journal Article
- Keywords:
Plasmodium falciparum;
Imported;
Magnetic nanoparticles separation;
Real-time PCR;
Detection
- From:
Chinese Journal of Schistosomiasis Control
2014;(5):522-525,530
- CountryChina
- Language:Chinese
-
Abstract:
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.
