Sanches Letícia da Cruz, Martini Cleber Costa de, Nakamura Alex Akira, Santiago Maria Emília Bodini, Dolabela de Lima Beatriz, Lima Valéria Marçal Felix de
Departamento de Clínica, Cirurgia e Reprodução Animal, Faculdade de Medicina Veterinária de Araçatuba, Universidade Estadual Paulista - UNESP, Araçatuba, SP, Brasil.
Zoológico Municipal de Bauru, Bauru, SP, Brasil.
Rev Bras Parasitol Vet. 2016 Oct-Dec;25(4):465-469. doi: 10.1590/S1984-29612016071. Epub 2016 Dec 1.
Leishmaniasis is a major public health problem worldwide. Because Leishmania can adapt to new hosts or vectors, knowledge concerning the current etiological agent in dogs is important in endemic areas. This study aimed to identify the Leishmania species detected in 103 samples of peripheral blood from dogs that were naturally infected with these protozoa. The diagnosis of leishmaniasis was determined through parasitological examination, the indirect enzyme-linked immunosorbent assay (ELISA) and the polymerase chain reaction (PCR). The Leishmania species were identified by means of PCR-restriction fragment length polymorphism (PCR-RFLP). The samples were subjected to PCR using oligonucleotide primers that amplify the intergenic region ITS1 of the rRNA gene in order to identify the species. The amplified DNA was digested using the restriction enzyme HaeIII. A restriction profile identical to L. amazonensis was shown in 77/103 samples and the profile was similar to L. infantum in 17/103. However, a mixed profile was shown in 9/103 samples, which impeded species identification. In conclusion, the infection in these dogs was predominantly due to L. amazonensis, thus indicating that diagnosing of cases of canine leishmaniasis needs to be reexamined, since the causative agent identified is not restricted to L. infantum.
利什曼病是全球主要的公共卫生问题。由于利什曼原虫能够适应新的宿主或媒介，在流行地区了解犬类当前的病原体情况非常重要。本研究旨在鉴定从自然感染这些原生动物的犬只的103份外周血样本中检测到的利什曼原虫种类。利什曼病的诊断通过寄生虫学检查、间接酶联免疫吸附测定（ELISA）和聚合酶链反应（PCR）来确定。利什曼原虫种类通过PCR-限制性片段长度多态性（PCR-RFLP）进行鉴定。使用寡核苷酸引物对样本进行PCR，扩增rRNA基因的ITS1基因间隔区以鉴定种类。扩增的DNA用限制性内切酶HaeIII进行消化。103份样本中有77份显示出与亚马逊利什曼原虫相同的限制性图谱，17份与婴儿利什曼原虫的图谱相似。然而，103份样本中有9份显示出混合图谱，这妨碍了种类鉴定。总之，这些犬只的感染主要是由亚马逊利什曼原虫引起的，因此表明犬类利什曼病病例的诊断需要重新审视，因为所确定的病原体并不局限于婴儿利什曼原虫。
