Reifenberg J M, Solano P, Duvallet G, Cuisance D, Simpore J, Cuny G
CIRAD-EMVT (Centre de coopération Internationale en Recherche Agronomique pour le Développement--Département d'Elevage et de Médecine Vétérinaire), Montpellier, France.
Vet Parasitol. 1997 Aug;71(4):251-62. doi: 10.1016/s0304-4017(97)00011-3.
A total of 33 trypanosome cryostabilates isolated from domestic animals (bovine and dogs) were analysed using the polymerase chain reaction (PCR). The PCR was undertaken on diluted and treated buffy coat solutions according to an easy protocol of purification, using primers specific to Trypanosoma (Nannomonas) congolense of Savannah, Riverine-Forest, Kilifi and Tsavo types, T. (N) simiae, T. (Trypanozoon) brucei and T. (Duttonella) vivax. The results showed a lack of PCR sensitivity when target solutions were simply diluted, probably a reflection of the inaccuracy of the dilution procedure at very low trypanosome numbers. Nine mixed infections were found in purified samples whereas only three were detected in diluted crude solutions. T. congolense Savannah-type was present in all stabilates. Double infections involving this type with the Riverine-Forest type, T. vivax or T. brucei, were found. One stabilate was found to be infected with the three trypanosome types, namely T. congolense Savannah and Riverine-Forest genotypes and T. vivax. No infection attributable to T. congolense Kilifi and Tsavo types or T. simiae was detected in these stabilates. This work confirmed the abundance of mixed infections in the field, which could not have been detected by the classical parasitological methods. Amongst the T. congolense infections, the Savannah genotype was found to be predominant over the Riverine-Forest type; that could be a consequence of differences in genotype virulence in cattle. The detection of T. congolense Riverine-Forest type in vertebrate hosts living in wet areas could be confirmation of the suspected affinity of relationships between this taxa and the riverine forest tsetse fly species.
使用聚合酶链反应（PCR）对从家畜（牛和狗）中分离出的33种锥虫冷冻稳定株进行了分析。根据一种简单的纯化方案，对稀释和处理后的血沉棕黄层溶液进行PCR，使用针对萨凡纳型、河滨森林型、基利菲型和察沃型的刚果锥虫（南诺莫纳锥虫）、猕猴锥虫、布氏锥虫（锥虫属）和活泼锥虫（达顿锥虫属）的特异性引物。结果表明，当靶溶液仅进行稀释时，PCR缺乏敏感性，这可能反映了在锥虫数量非常低时稀释程序的不准确性。在纯化样品中发现了9例混合感染，而在稀释的粗溶液中仅检测到3例。所有稳定株中均存在萨凡纳型刚果锥虫。发现了该类型与河滨森林型、活泼锥虫或布氏锥虫的双重感染。发现一个稳定株感染了三种锥虫类型，即萨凡纳型和河滨森林型刚果锥虫以及活泼锥虫。在这些稳定株中未检测到归因于基利菲型和察沃型刚果锥虫或猕猴锥虫的感染。这项工作证实了野外混合感染的普遍性，而这是传统寄生虫学方法无法检测到的。在刚果锥虫感染中，发现萨凡纳基因型比河滨森林型更占优势；这可能是牛群中基因型毒力差异的结果。在生活在湿润地区的脊椎动物宿主中检测到河滨森林型刚果锥虫，这可能证实了该分类群与河滨森林采采蝇物种之间疑似的亲缘关系。
