PET-PCR method for the molecular detection of malaria parasites in a national malaria surveillance study in Haiti, 2011
| Autor: | Dragan Ljolje, Eniko Edit Akom, Naomi W. Lucchi, Curtis S. Huber, Samuel E. Jean, Mara A. Karell, Roland Oscar, Ito Journel, Josiane Buteau, Venkatachalam Udhayakumar, Jacques Boncy, John W. Barnwell, Eric Rogier, Kimberly E. Mace, Ira F. Goldman |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Biology
Polymerase Chain Reaction law.invention law Diagnosis medicine TaqMan Humans Mass Screening Polymerase chain reaction Mass screening Research Plasmodium falciparum DNA Protozoan medicine.disease biology.organism_classification Virology Haiti Malaria PET-PCR Infectious Diseases Blood Parasitology Molecular Diagnostic Techniques Epidemiological Monitoring Pcr method Nested polymerase chain reaction |
| Zdroj: | Malaria Journal |
| ISSN: | 1475-2875 |
| Popis: | Background: Recently, a real-time PCR assay known as photo-induced electron transfer (PET)-PCR which relies on self-quenching primers for the detection of Plasmodium spp. and Plasmodium falciparum was described. PET-PCR assay was found to be robust, and easier to use when compared to currently available real-time PCR methods. The potential of PET-PCR for molecular detection of malaria parasites in a nationwide malaria community survey in Haiti was investigated. Methods: DNA from the dried blood spots was extracted using QIAGEN methodology. All 2,989 samples were screened using the PET-PCR assay in duplicate. Samples with a cycle threshold (CT) of 40 or less were scored as positive. A subset of the total samples (534) was retested using a nested PCR assay for confirmation. In addition, these same samples were also tested using a TaqMan-based real-time PCR assay. Results: A total of 12 out of the 2,989 samples screened (0.4%) were found to be positive by PET-PCR (mean CT value of 35.7). These same samples were also found to be positive by the nested and TaqMan-based methods. The nested PCR detected an additional positive sample in a subset of 534 samples that was not detected by either PET-PCR or TaqMan-based PCR method. Conclusion: While the nested PCR was found to be slightly more sensitive than the PET-PCR, it is not ideal for high throughput screening of samples. Given the ease of use and lower cost than the nested PCR, the PET-PCR provides an alternative assay for the rapid screening of a large number of samples in laboratory settings. |
| Databáze: | OpenAIRE |
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