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Title: Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys
Authors: Peipei Li
Zhenjun Zhao
Ying Wang
Hua Xing
Daniel M. Parker
Zhaoqing Yang
Elizabeth Baum
Wenli Li
Jetsumon Sattabongkot
Jeeraphat Sirichaisinthop
Shuying Li
Guiyun Yan
Liwang Cui
Qi Fan
Dalian Institute of Biotechnology
Third Military Medical University
Dalian University of Technology
Pennsylvania State University
Kunming Medical University
University of California, Irvine
Mahidol University
Vector Borne Disease Training Center
Keywords: Immunology and Microbiology;Medicine
Issue Date: 8-May-2014
Citation: Malaria Journal. Vol.13, No.1 (2014)
Abstract: Background: Nested PCR is considered a sensitive and specific method for detecting malaria parasites and is especially useful in epidemiological surveys. However, the preparation of DNA templates for PCR is often time-consuming and costly. Methods. A simplified PCR method was developed to directly use a small blood filter paper square (2 × 2 mm) as the DNA template after treatment with saponin. This filter paper-based nested PCR method (FP-PCR) was compared to microscopy and standard nested PCR with DNA extracted by using a Qiagen DNA mini kit from filter paper blood spots of 204 febrile cases. The FP-PCR technique was further applied to evaluate malaria infections in 1,708 participants from cross-sectional epidemiological surveys conducted in Myanmar and Thailand. Results: The FP-PCR method had a detection limit of ∼0.2 parasites/μL blood, estimated using cultured Plasmodium falciparum parasites. With 204 field samples, the sensitivity of the FP-PCR method was comparable to that of the standard nested PCR method, which was significantly higher than that of microscopy. Application of the FP-PCR method in large cross-sectional studies conducted in Myanmar and Thailand detected 1.9% (12/638) and 6.2% (66/1,070) asymptomatic Plasmodium infections, respectively, as compared to the detection rates of 1.3% (8/638) and 0.04% (4/1,070) by microscopy. Conclusion: This FP-PCR method was much more sensitive than microscopy in detecting Plasmodium infections. It drastically increased the detection sensitivity of asymptomatic infections in cross-sectional surveys conducted in Thailand and Myanmar, suggesting that this FP-PCR method has a potential for future applications in malaria epidemiology studies. © 2014 Li et al.; licensee BioMed Central Ltd.
ISSN: 14752875
Appears in Collections:Scopus 2011-2015

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