Monitoring and evaluation of lymphatic filariasis interventions: an improved PCR-based pool screening method for high throughput Wuchereria bancrofti detection using dried blood spots

Particle and Fibre Toxicology

Aurore Lemoine¹  Catherine Plichart¹ 

[1] Institut Louis Malardé, Papeete, Tahiti, French Polynesia

关键词: French Polynesia;    Blood spots;    Pool screen;    Real-time PCR;    Disease monitoring;    Lymphatic filariasis;   

Others  :  1227992
DOI  :  10.1186/1756-3305-6-110

 received in 2012-12-09, accepted in 2013-04-14,  发布年份 2013

【 摘 要 】

Background

Effective diagnostic tools are necessary to monitor and evaluate interruption of Lymphatic Filariasis (LF) transmission. Accurate detection of Wuchereria bancrofti (Wb) microfilaria (mf) is essential to measure the impact of community treatment programmes. PCR-based assays are specific, highly sensitive tools allowing the detection of Wuchereria bancrofti DNA in human blood samples. However, current protocols describing the pool screening approach, use samples of less than 60 μl of blood, which limits the sensitivity of the pool-screen PCR assay. The purpose of this study was to improve the pool-screen PCR protocol to enhance its sensitivity and usefulness for population scale studies.

Findings

DNA extractions were performed with the DNeasy kit, the PCR with the Wb LDR primers and the SYBR-Green dye. Improvements of our pool-screen real-time PCR (qPCR) assay allowed the detection of as little as one Wb microfilaria diluted in a pool of at least 12 blood samples of 60 μl each. Using this assay, mf burdens can be predicted using a standard curve derived from mf spiked dried blood samples. The sensitivity achieved is equivalent to the detection of a single LF positive individual carrying a mf burden as low as 18 mf/ml, in a pool of blood samples from at least 12 individuals.

Conclusions

Due to its sensitivity, rapidity and cost-effectiveness, we suggest this qPCR pool-screening assay could be used as a diagnostic tool for population- scale filariasis elimination monitoring and evaluation.

【 授权许可】

   
2013 Plichart and Lemoine; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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