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. 2014 Nov 26:13:462.
doi: 10.1186/1475-2875-13-462.

PET-PCR method for the molecular detection of malaria parasites in a national malaria surveillance study in Haiti, 2011

Naomi W Lucchi  1 , Mara A Karell, Ito Journel, Eric Rogier, Ira Goldman, Dragan Ljolje, Curtis Huber, Kimberly E Mace, Samuel E Jean, Eniko E Akom, Roland Oscar, Josiane Buteau, Jacques Boncy, John W Barnwell, Venkatachalam Udhayakumar
Affiliations

PET-PCR method for the molecular detection of malaria parasites in a national malaria surveillance study in Haiti, 2011

Naomi W Lucchi et al. Malar J. .

Abstract

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.

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Figures

Figure 1
Figure 1
Sample processing.
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