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

Malaria Journal

Research

Dragan Ljolje¹  Mara A Karell¹  Eric Rogier²  Ira Goldman²  Curtis Huber²  John W Barnwell²  Kimberly E Mace²  Naomi W Lucchi²  Venkatachalam Udhayakumar²  Ito Journel³  Josiane Buteau³  Jacques Boncy³  Eniko E Akom⁴  Samuel E Jean⁴  Roland Oscar⁵ 

[1] Atlanta Research and Education Foundation, Decatur, GA, USA;Centers for Disease Control and Prevention, Center for Global Health, Division of Parasitic Diseases and Malaria, Malaria Branch, Atlanta, GA, USA;Laboratoire National de Santé Publique, Port au Prince, Haiti;Population Services International-Haiti, Port au Prince, Haiti;Programme National de Contrôle de la Malaria, Port au Prince, Haiti;

关键词: Malaria;    Diagnosis;    Haiti;    PET-PCR;   

DOI  :  10.1186/1475-2875-13-462

 received in 2014-08-19, accepted in 2014-11-19,  发布年份 2014

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© Lucchi et al.; licensee BioMed Central Ltd. 2014