Molecular Diagnosis of Malaria by Photo-Induced Electron Transfer Fluorogenic Primers: PET-PCR
| Autor: | Alexandre J. DaSilva, Simon Kariuki, Mara A. Karell, Vincent R. Hill, Jothikumar Narayanan, Venkatachalam Udhayakumar, Maniphet V. Xayavong, Naomi W. Lucchi |
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| Rok vydání: | 2013 |
| Předmět: |
Epidemiology
Plasmodium falciparum lcsh:Medicine Parasitemia Protozoology Biology Global Health Polymerase Chain Reaction Sensitivity and Specificity Microbiology law.invention Diagnostic Medicine law parasitic diseases RNA Ribosomal 18S Parasitic Diseases medicine Humans lcsh:Science Epidemiological Methods Polymerase chain reaction DNA Primers Fluorescent Dyes Multidisciplinary Population Biology lcsh:R Tropical Diseases (Non-Neglected) Gold standard (test) medicine.disease biology.organism_classification Virology Malaria Diagnosis of malaria Infectious Diseases Parastic Protozoans Medicine lcsh:Q Primer (molecular biology) Nested polymerase chain reaction Research Article |
| Zdroj: | PLoS ONE, Vol 8, Iss 2, p e56677 (2013) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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