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Parasitology Research

, Volume 106, Issue 3, pp 621–625 | Cite as

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

  • Tongjit Thanchomnang
  • Pewpan M. IntapanEmail author
  • Sudchit Chungpivat
  • Viraphong Lulitanond
  • Wanchai Maleewong
Original Paper

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.

Keywords

Fluorescence Resonance Energy Transfer Melting Curve Analysis Babesia Differential Detection Restriction Fragment Length Polymorphism Profile 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

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.

Conflict of interest statement

No conflict of interest is declared.

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Tongjit Thanchomnang
    • 1
  • Pewpan M. Intapan
    • 1
    Email author
  • Sudchit Chungpivat
    • 3
  • Viraphong Lulitanond
    • 2
  • Wanchai Maleewong
    • 1
  1. 1.Department of Parasitology, Faculty of MedicineKhon Kaen UniversityKhon KaenThailand
  2. 2.Department of Microbiology, Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious DiseasesKhon Kaen UniversityKhon KaenThailand
  3. 3.Parasitology Unit, Department of Pathology, Faculty of Veterinary ScienceChulalongkorn UniversityBangkokThailand

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