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

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

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.

Original languageEnglish (US)
Article number175
JournalMalaria journal
Volume13
Issue number1
DOIs
StatePublished - May 8 2014

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Infectious Diseases

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