Manterola Carlos, Benavente Felipe, Melo Angélica, Vial Manuel, Roa Juan Carlos
Department of Surgery, Faculty of Medicine, Universidad de La Frontera, Temuco, Chile.
Parasitol Int. 2008 Sep;57(3):342-6. doi: 10.1016/j.parint.2008.02.005. Epub 2008 Mar 4.
Echinococcus granulosus species has a wide variety in both geography and hosts; indeed, 10 genotypes have been reported in studies on material of animal origin. The aim of this study was to genotype E. granulosus obtained from human hydatid cysts.
The hydatid fluid and sand was collected from patients who underwent surgery for hepatic and pulmonary hydatidosis at Hospital Regional in Temuco, Chile, between 2004 and 2005. Two PCR systems were used: PCR Eg 9 and PCR Eg 16. The RsaI enzyme was used for RFLP. The genotype was confirmed using the sequence of one fragment of 366 bp from a mitochondrial gene (cox1).
The DNA of protoscolices from 24 samples was analyzed, 4 of them from pulmonary cysts and 20 from hepatic cysts. The 366 bp fragment was amplified in 20 out of 24 samples (83.3%). Enzymatic digestion revealed the presence of 3 possible genotypes: in 20 out of 21 samples (95,2%), a restriction was observed corresponding to the G1 or G7 genotypes; in the remaining sample genotype G4 or G7 was observed. Sequencing confirmed the presence of G1 genotype for 19 samples and G6 genotype for the remaining sample (G4 or G7 according to PCR-RFLP).
The PCR-RFLP technique enabled three possible genotypes present (G1 or G7, G4 or G7) to be established. Sequencing allowed us to decisively identify the G1 and G6 genotypes in our study group. Previous studies agree with the identification of the G1 genotype in our country. We consider it significant that the G6 genotype is present in Chile for its epidemiological implications.
细粒棘球绦虫在地理分布和宿主方面具有广泛的多样性;事实上,在对动物源材料的研究中已报道了10种基因型。本研究的目的是对从人包虫囊肿中获得的细粒棘球绦虫进行基因分型。
2004年至2005年期间,从智利特木科地区医院接受肝和肺包虫病手术的患者中收集包虫液和包虫砂。使用了两种PCR系统:PCR Eg 9和PCR Eg 16。使用RsaI酶进行限制性片段长度多态性分析(RFLP)。通过线粒体基因(cox1)的一个366 bp片段的序列确认基因型。
分析了24个样本中原头蚴的DNA,其中4个来自肺囊肿,20个来自肝囊肿。24个样本中有20个(83.3%)扩增出了366 bp片段。酶切显示存在3种可能的基因型:21个样本中有20个(95.2%)观察到与G1或G7基因型相对应的限制性酶切;在其余样本中观察到G4或G7基因型。测序确认19个样本存在G1基因型,其余样本存在G6基因型(根据PCR-RFLP为G4或G7)。
PCR-RFLP技术能够确定存在的3种可能基因型(G1或G7、G4或G7)。测序使我们能够在研究组中明确鉴定出G1和G6基因型。先前的研究与我国对G1基因型的鉴定结果一致。我们认为G6基因型在智利的存在因其流行病学意义而具有重要意义。