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Monitoring and evaluation of lymphatic filariasis interventions: an improved PCR-based pool screening method for high throughput Wuchereria bancrofti detection using dried blood spots
- Source :
- Parasites & Vectors
- Publication Year :
- 2013
- Publisher :
- Springer Science and Business Media LLC, 2013.
-
Abstract
- 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.
- Subjects :
- Veterinary medicine
Time Factors
Cost-Benefit Analysis
Population
Short Report
Blood spots
French Polynesia
Biology
Real-Time Polymerase Chain Reaction
medicine.disease_cause
Sensitivity and Specificity
Microfilaria
Filariasis
Elephantiasis, Filarial
medicine
Animals
Humans
Wuchereria bancrofti
education
Microfilariae
Lymphatic filariasis
DNA Primers
Disease monitoring
education.field_of_study
DNA, Helminth
Reference Standards
medicine.disease
Virology
Standard curve
Infectious Diseases
Real-time polymerase chain reaction
Parasitology
Pool screen
Real-time PCR
Subjects
Details
- ISSN :
- 17563305
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Parasites & Vectors
- Accession number :
- edsair.doi.dedup.....a2fb44ebeae8cc291241b319be10fb7c