Skotarczak Bogumiła
Katedra Genetyki, Uniwersytet Szczeciński, Al. Piastów 40B, 71-065 Szczecin.
Wiad Parazytol. 2007;53(4):271-80.
This article presents the current state of our knowledge on babesiosis (piroplasmosis), one of the dangerous, invasive disease of humans and animals, transmitted by ticks. It is included among emerging diseases because its spread and significance have increased in recent years. This sickness is caused by intraerythrocytic parasites belonging to the Babesia species and it is a well-known zoonosis occurring in animals; as a human disease it was unknown almost till the first half of the last century. The intensified migration of human population and human interference in a forest biotope caused that number of recognized cases has grown considerably in recent years. Piroplasmosis in dogs is widely spread all over the world and it is caused by several Babesia species. The principal etiological factor of babesiosis in dogs is B. canis, which turned out to be a collective species represented by three subspecies for which the vectors are three different species of ticks. Their geographical extent indicates the endemic areas for this often fatal disease. A technique, the most often applied in the detection of Babesia is a full blood smear stained with Giemsa or Wright method. However, the estimation of the specimen depends to a large extent on the experience of the diagnostician. The immunological and serological methods are characterized with a high specificity and sensitivity but there are patients in which the false negative results have been obtained. Therefore, the traditional methods have been complemented or even ousted by the molecular methods, in which polymerase chain reaction (PCR) brings the biggest profits. However, the standardization of this technique still remains under elaboration. The usefulness of the PCR protocol has been tested with different molecular destinations from which sequences of genes encoding rRNA for small ribosomal subunit are taken into consideration. Within ribosome, the evolutionally conservative areas can be distinguished, i.e. having the nucleotide sequences similar to the majority or all Babesia species and to others closely related to them. Such construction of gene enables designing of starters complementary to conservative sites to PCR, detecting a large group of related organisms. Another molecular marker allowing on the accurate identification of Babesia is gene encoding the beta-tubuline protein. There are two introns within this gene, from which the first one shows a big variability with regard to the length as well as to the nucleotide sequence, therefore, the PCR products show a diverse length depending on the Babesia species. But these differences are too small for some species and, confirming methods that extend time of diagnostics are essential. The other genes which sequences can be used as molecular aim to the detection of DNA and Babesia species diversification are genes encoding the Heat Shock Proteins HSP 70. However, the gene hsp 70 shows a big conservatism of the nucleotide sequence even between the non related organisms; therefore, this method, based on the amplification of whole genome or its fragments, applies mainly in analysis of molecular phylogenetic.
本文介绍了我们目前对巴贝斯虫病(梨形虫病)的认识,它是一种由蜱传播的、对人类和动物都有危害的侵袭性疾病。它被列入新兴疾病,因为近年来其传播范围和重要性都有所增加。这种疾病由属于巴贝斯虫属的红细胞内寄生虫引起,是一种在动物中广为人知的人畜共患病;作为一种人类疾病,直到上世纪上半叶几乎还不为人所知。人类人口迁移的加剧以及人类对森林生物群落的干扰导致近年来确诊病例数量大幅增加。犬梨形虫病在世界各地广泛传播,由几种巴贝斯虫引起。犬巴贝斯虫病的主要病因是犬巴贝斯虫,它原来是一个由三个亚种代表的集合物种,其传播媒介是三种不同的蜱。它们的地理分布表明了这种常致命疾病的流行区域。检测巴贝斯虫最常用的技术是用吉姆萨或瑞氏方法染色的全血涂片。然而,对样本的评估在很大程度上取决于诊断医生的经验。免疫和血清学方法具有高特异性和高敏感性,但有些患者会出现假阴性结果。因此,传统方法已被分子方法补充甚至取代,其中聚合酶链反应(PCR)带来的收益最大。然而,这项技术的标准化仍在完善中。已用不同的分子目标测试了PCR方案的有效性,其中考虑了从小核糖体亚基编码rRNA的基因序列。在核糖体中,可以区分进化上保守的区域,即具有与大多数或所有巴贝斯虫物种以及与其密切相关的其他物种相似的核苷酸序列。这种基因结构使得能够设计与PCR保守位点互补的引物,从而检测一大群相关生物体。另一个能够准确鉴定巴贝斯虫的分子标记是编码β -微管蛋白的基因。该基因内有两个内含子,其中第一个内含子在长度和核苷酸序列方面表现出很大的变异性,因此,PCR产物的长度因巴贝斯虫物种而异。但对于某些物种来说,这些差异太小,因此,延长诊断时间的确认方法至关重要。其他可将其序列用作检测DNA和巴贝斯虫物种多样性的分子目标的基因是编码热休克蛋白HSP 70的基因。然而,hsp 70基因即使在非相关生物体之间也表现出核苷酸序列的高度保守性;因此,这种基于全基因组或其片段扩增的方法主要应用于分子系统发育分析。