Cox Andrew, Tilley Aimee, McOdimba Francis, Fyfe Jenna, Eisler Mark, Hide Geoff, Welburn Susan
Centre for Tropical Veterinary Medicine, Royal (Dick) School of Veterinary Medicine, University of Edinburgh, Easter Bush, Roslin, Midlothian EH25 9RG, Scotland, UK.
Exp Parasitol. 2005 Sep;111(1):24-9. doi: 10.1016/j.exppara.2005.03.014.
Direct PCR analysis of trypanosome infected blood samples in the quantities required for large scale epidemiological study has always been problematic. Current methods for identifying and differentiating trypanosomes typically require several species-specific reactions, many of which rely on mouse passaged samples to obtain quality concentrated genomic DNA. As a consequence important epidemiological information may be lost during the sample preparation stage. Here, we report a PCR methodology that reduces processing and improves on the sensitivity of present screening methods. The PCR technique targets the gene encoding the small ribosomal subunit in order to identify and differentiate all clinically important African trypanosome species and some subspecies. The method is more economical, simple, and sensitive than current screening methods, and yields more detailed information, thereby making it a viable tool for large-scale epidemiological studies.
对大规模流行病学研究所需数量的锥虫感染血液样本进行直接PCR分析一直存在问题。目前用于鉴定和区分锥虫的方法通常需要几种物种特异性反应，其中许多反应依赖于经小鼠传代的样本以获得高质量的浓缩基因组DNA。因此，重要的流行病学信息可能在样本制备阶段丢失。在此，我们报告一种PCR方法，该方法减少了处理步骤并提高了现有筛查方法的灵敏度。该PCR技术针对编码小核糖体亚基的基因，以鉴定和区分所有具有临床重要性的非洲锥虫物种和一些亚种。该方法比目前的筛查方法更经济、简单且灵敏，并能产生更详细的信息，从而使其成为大规模流行病学研究的可行工具。
