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.
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.
Thanchomnang
Tongjit
T
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Intapan
Pewpan M
PM
Chungpivat
Sudchit
S
Lulitanond
Viraphong
V
Maleewong
Wanchai
W
D000818:Animals; D001769:Blood / Q000469:parasitology; D017178:Brugia malayi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017179:Brugia pahangi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology; D004285:Dogs; D005368:Filariasis / Q000175:diagnosis / Q000662:veterinary*; D031541:Fluorescence Resonance Energy Transfer; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D044366:Transition Temperature
D017931:DNA Primers; D015345:Oligonucleotide Probes