Abstract
BackgroundRecently, 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.MethodsDNA 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.ResultsA 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.ConclusionWhile 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.
Highlights
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
The prevalence and distribution of malaria infection in a country is an integral indicator for national malaria control programmes, which can be used to measure the success of intervention strategies and for appropriate resource allocation
Survey and specimen collection In November-December 2011, during peak malaria transmission season, a national population-based community survey was conducted by Population Services International (PSI), Haiti, in collaboration with the Haitian national malaria control programme (PNCM, per its acronym in French), and the National Public Health Laboratory (LNSP, per its acronym in French)
Summary
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. Studies have demonstrated that asymptomatic malaria infections can serve as transmission foci in both low and high transmission settings and, remain as sources of new infections [9,10,11,12,13]. It is imperative for malaria elimination programmes to detect as many malaria parasite infections as is possible, whether symptomatic or asymptomatic [7]
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