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.
journal_name
Malar Jjournal_title
Malaria journalauthors
Lucchi NW,Karell MA,Journel I,Rogier E,Goldman I,Ljolje D,Huber C,Mace KE,Jean SE,Akom EE,Oscar R,Buteau J,Boncy J,Barnwell JW,Udhayakumar Vdoi
10.1186/1475-2875-13-462subject
Has Abstractpub_date
2014-11-26 00:00:00pages
462issn
1475-2875pii
1475-2875-13-462journal_volume
13pub_type
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