Molecular Diagnosis of Malaria by Photo-Induced Electron Transfer Fluorogenic Primers: PET-PCR

Details

• Alternative Title:
  PLoS One
• Personal Author:
  Lucchi, Naomi W. ; Narayanan, Jothikumar ; Karell, Mara A. ; Xayavong, Maniphet ; Kariuki, Simon ; DaSilva, Alexandre J. ; Hill, Vincent ; Udhayakumar, Venkatachalam
• Description:
  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.
• Subjects:
  DNA Primers Fluorescent Dyes Humans Malaria Plasmodium Falciparum Polymerase Chain Reaction RNA, Ribosomal, 18S Sensitivity And Specificity
• Source:
  PLoS One. 2013; 8(2).
• Document Type:
  Journal Article
• Volume:
  8
• Issue:
  2
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:e024cc08e3f3c326b8d19137f9065751dc9222f60df51dd5fe2ad256afd9e57a
• Download URL:
  https://stacks.cdc.gov/view/cdc/21654/cdc_21654_DS1.pdf
• File Type:
  [PDF - 247.39 KB ]