Abstract
A LightCycler real-time polymerase chain reaction (PCR) assay was developed to detect Wuchereria bancrofti DNA in blood-fed mosquitoes. The assay is based on fluorescence melting curve analysis of the PCR product generated from a family of repeated DNA elements: the 182 bp SspI repeat, specific to the genus Wuchereria. According to the melting temperature, W. bancrofti infected-mosquitoes were differentiated from Brugia malayi-infected and non-infected mosquitoes as well as from genomic DNA of Dirofilaria immitis and human DNA. The method proved to be 100% sensitive in all W. bancrofti-infected mosquitoes. Melting curve analysis offers a rapid alternative for the specific detection of W. bancrofti in mosquitoes. It is very accurate and sensitive, allows a high throughput and can be performed on very small samples. The method therefore has great potential for application in epidemiological studies.
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Acknowledgements
This study was supported by the Diagnostic and Research Center of Emerging and Re-emerging Infectious Diseases of Northeastern Thailand and the Faculty of Medicine, Khon Kaen University. The authors wish to thank Dr. Mark Roselieb for his assistance in manuscript preparation. The animal experiments used in this study comply with the current Thai laws.
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Lulitanond, V., Intapan, P.M., Pipitgool, V. et al. Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and melting curve analysis. Parasitol Res 94, 337–341 (2004). https://doi.org/10.1007/s00436-004-1221-6
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DOI: https://doi.org/10.1007/s00436-004-1221-6