Abstract
A real-time fluorescence resonance energy transfer PCR combined with melting curve analysis was developed for differentiating Brugia malayi and Brugia pahangi DNA in host blood using one set of primers and fluorophore-labeled hybridization probes specific for HhaI repetitive DNA. The differentiation of both species was based on their melting temperatures (Tm). The mean Tm ± SD of B. malayi and B. pahangi were 56.18 ± 0.21 and 52.49 ± 0.07, respectively. The method was used for the molecular detection of B. pahangi in infected dog blood samples. The diagnostic sensitivity, specificity, accuracy, and positive and negative predictive values of this method were 100%. The detected mean difference of the Tm might allow the simple discrimination of two related species. This method is fast, sensitive, allows for a high throughput, can be performed on very small volumes, and has potential for diagnosis of B. pahangi-infected dogs in endemic areas as well as for large epidemiological investigations.
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Acknowledgments
This study was supported by grants from the Office of the Higher Education Commission (grant funded under the Strategic Scholarships for Frontier Research Networks), the Thailand Research Fund, and the Research and Technology Transfer Affairs and the Faculty of Medicine (research grant Comm51201), Khon Kaen University, Thailand. The authors wish to thank Wej Choochote for his technical support. All investigations in this study complied with current Thai laws.
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Thanchomnang, T., Intapan, P.M., Chungpivat, S. et al. Differential detection of Brugia malayi and Brugia pahangi by real-time fluorescence resonance energy transfer PCR and its evaluation for diagnosis of B. pahangi-infected dogs. Parasitol Res 106, 621–625 (2010). https://doi.org/10.1007/s00436-009-1706-4
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DOI: https://doi.org/10.1007/s00436-009-1706-4