title: Molecular diagnosis of malaria by photo-induced electron transfer fluorogenic primers: PET-PCR.
abstract: There is a critical need for developing new malaria diagnostic tools that are sensitive, cost effective and capable of performing large scale diagnosis. The real-time PCR methods are particularly robust for large scale screening and they can be used in malaria control and elimination programs. We have designed novel self-quenching photo-induced electron transfer (PET) fluorogenic primers for the detection of P. falciparum and the Plasmodium genus by real-time PCR. A total of 119 samples consisting of different malaria species and mixed infections were used to test the utility of the novel PET-PCR primers in the diagnosis of clinical samples. The sensitivity and specificity were calculated using a nested PCR as the gold standard and the novel primer sets demonstrated 100% sensitivity and specificity. The limits of detection for P. falciparum was shown to be 3.2 parasites/µl using both Plasmodium genus and P. falciparum-specific primers and 5.8 parasites/µl for P. ovale, 3.5 parasites/µl for P. malariae and 5 parasites/µl for P. vivax using the genus specific primer set. Moreover, the reaction can be duplexed to detect both Plasmodium spp. and P. falciparum in a single reaction. The PET-PCR assay does not require internal probes or intercalating dyes which makes it convenient to use and less expensive than other real-time PCR diagnostic formats. Further validation of this technique in the field will help to assess its utility for large scale screening in malaria control and elimination programs.
authors: Division of Parasitic Diseases and Malaria, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, USA. NLucchi@cdc.gov
authors: Naomi W
authors: NW
authors: Lucchi
authors: Jothikumar
authors: J
authors: Narayanan
authors: Mara A
authors: MA
authors: Karell
authors: Maniphet
authors: M
authors: Xayavong
authors: Simon
authors: S
authors: Kariuki
authors: Alexandre J
authors: AJ
authors: DaSilva
authors: Vincent
authors: V
authors: Hill
authors: Venkatachalam
authors: V
authors: Udhayakumar
mesh_terms: D017931:DNA Primers; D005456:Fluorescent Dyes; D006801:Humans; D008288:Malaria / Q000175:diagnosis* / Q000235:genetics / Q000469:parasitology; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D012337:RNA, Ribosomal, 18S