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A Novel qPCR Assay for the Detection of African Animal Trypanosomosis in Trypanotolerant and Trypanosusceptible Cattle Breeds

机译:新型qPCR检测非洲锥虫病和锥虫易感牛品种中的锥虫病的检测

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摘要

This study was conducted to (i) determine the prevalence of African Animal Trypanosomosis (AAT) in tsetse challenged areas, (ii) compare conventional with qPCR detection systems and (iii) evaluate the host genetic background and biology as risk factors. AAT prevalence studies are often confronted with low levels of parasitaemia. Hence, we designed a novel qPCR assay using primers and species specific probes amplifying the Internal Transcribed Spacer 1 (ITS1) gene. Thereby all three AAT species could be detected simultaneously. 368 individuals from three cattle types (Baoulé, Zebu and hybrids) originating from 72 farms in Burkina Faso were analysed. Farmers were interviewed and morphometric measurements of the cattle taken. A chi-squared test and a logistic regression model were calculated to detect associations with infection. In our study, the overall rate of prevalence detected with the novel qPCR assay was 11.14%. Compared to conventional PCR we identified a concordance of 91.30%. We tested 41 animals positive for trypanosome DNA, five animals showed multiple infections. Zebus were twice as often infected (21.74%) compared to Baoulé (9.70%) and hybrids (9.57%). Trypanosoma vivax is the dominant species (9.24%), as compared to T. congolense (2.44%) and T. brucei (0.82%). The chi-squared tests linking the infection events to the breeds (Zebu vs. Baoulé and Zebu vs. hybrids) were on the border of significance. No significant association with other tested parameters could be detected. We introduce a novel qPCR technique for the fast, sensitive and simultaneous detection of the three AAT species. Our results suggest that associations with breed and infection exist since Zebu cattle are more likely to be infected compared to Baoulé and hybrids. Indigenous taurine cattle breeds, like the Baoulé, therefore provide a unique and valuable genetic resource.
机译:进行这项研究的目的是:(i)确定采采蝇感染地区的非洲动物锥虫病(AAT)的患病率;(ii)将常规与qPCR检测系统进行比较;(iii)评价宿主的遗传背景和生物学作为危险因素。 AAT患病率研究通常面临低水平的寄生虫血症。因此,我们使用引物和物种特异性探针扩增内部转录间隔子1(ITS1)基因设计了一种新颖的qPCR分析方法。因此,可以同时检测所有三种AAT物质。分析了来自布基纳法索的72个农场的三种牛(鲍乌勒,泽布和杂种)的368个人。采访了农民,并对牛进行了形态测量。计算卡方检验和逻辑回归模型以检测与感染的关联。在我们的研究中,新型qPCR分析检测到的总患病率为11.14%。与常规PCR相比,我们确定了91.30%的一致性。我们测试了41只锥虫DNA阳性的动物,五只动物表现出多重感染。 Zebus的经常性感染(21.74%)是鲍勒(9.70%)和杂种(9.57%)的两倍。间虫锥虫是主要物种(9.24%),而锥虫(2.44%)和布氏锥虫(0.82%)是优势种。卡方检验将感染事件与该品种(Zebu与Baoulé和Zebu与杂种)联系起来。与其他测试参数没有显着关联。我们介绍了一种新颖的qPCR技术,用于快速,灵敏和同时检测三种AAT物种。我们的结果表明,与Ze牛和杂种相比,由于瘤牛牛更容易被感染,因此存在与繁殖和感染的关联。因此,像鲍勒(Baoulé)这样的土著牛磺酸牛品种提供了独特而宝贵的遗传资源。

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