Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys
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 u...
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th-mahidol.339702018-11-09T09:37:12Z Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys 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 Immunology and Microbiology Medicine 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. 2018-11-09T02:21:36Z 2018-11-09T02:21:36Z 2014-05-08 Article Malaria Journal. Vol.13, No.1 (2014) 10.1186/1475-2875-13-175 14752875 2-s2.0-84901030223 https://repository.li.mahidol.ac.th/handle/123456789/33970 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84901030223&origin=inward |
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Immunology and Microbiology Medicine 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 Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
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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. |
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Dalian Institute of Biotechnology |
author_facet |
Dalian Institute of Biotechnology 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 |
format |
Article |
author |
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 |
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Peipei Li |
title |
Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
title_short |
Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
title_full |
Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
title_fullStr |
Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
title_full_unstemmed |
Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys |
title_sort |
nested pcr detection of malaria directly using blood filter paper samples from epidemiological surveys |
publishDate |
2018 |
url |
https://repository.li.mahidol.ac.th/handle/123456789/33970 |
_version_ |
1763487237074321408 |