Title |
Molecular Diagnosis of Malaria by Photo-Induced Electron Transfer Fluorogenic Primers: PET-PCR
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Published in |
PLOS ONE, February 2013
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DOI | 10.1371/journal.pone.0056677 |
Pubmed ID | |
Authors |
Naomi W. Lucchi, Jothikumar Narayanan, Mara A. Karell, Maniphet Xayavong, Simon Kariuki, Alexandre J. DaSilva, Vincent Hill, Venkatachalam Udhayakumar |
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. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United Kingdom | 1 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United Kingdom | 1 | <1% |
Greece | 1 | <1% |
Estonia | 1 | <1% |
Brazil | 1 | <1% |
Unknown | 139 | 97% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 21 | 15% |
Student > Master | 21 | 15% |
Student > Bachelor | 17 | 12% |
Researcher | 16 | 11% |
Student > Postgraduate | 9 | 6% |
Other | 28 | 20% |
Unknown | 31 | 22% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 31 | 22% |
Agricultural and Biological Sciences | 26 | 18% |
Immunology and Microbiology | 13 | 9% |
Medicine and Dentistry | 13 | 9% |
Nursing and Health Professions | 9 | 6% |
Other | 14 | 10% |
Unknown | 37 | 26% |