Detection of mutant P2 adenosine transporter (TbAT1) gene in Trypanosoma brucei gambiense isolates from northwest Uganda using allele-specific polymerase chain reaction.
作者： Barbara Nerima , Enock Matovu , George W. Lubega , John C. K. Enyaru
DOI: 10.1111/J.1365-3156.2007.01918.X
关键词: Restriction fragment length polymorphism 、 Biology 、 Variants of PCR 、 Allele 、 Mutant 、 Polymerase chain reaction 、 Virology 、 Polymorphism (computer science) 、 Trypanosoma brucei 、 Melarsoprol
摘要: OBJECTIVE To assess the application of allele-specific PCR (AS-PCR) as a fast, cheap and reliable method for detecting mutant TbAT1 associated with melarsoprol relapse in Trypanosoma brucei gambiense isolates from northwest Uganda. METHODS A total 105 trypanosome were analysed using SfaN1 restriction fragment length polymorphism (RFLP) AS-PCR, former used gold standard. Sensitivity, specificity, positive negative predictive values AS-PCR well agreement between tests determined. RESULTS Eleven had while 94 exhibited wild-type genes. There was highly significant RFLP kappa intra-class correlation 1.0. The sensitivity specificity both 100%, found to be equal Cost time analyses performed 4.3 times cheaper than RFLP, addition less required its execution. CONCLUSION should test choice screening ever-increasing numbers field isolates.
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