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通过低严谨度单特异性引物聚合酶链反应揭示的从巴西东北部锥蝽分离出的克氏锥虫I的遗传多态性

Genetic polymorphism in Trypanosoma cruzi I isolated from Brazilian Northeast triatomines revealed by low-stringency single specific primer-polymerase chain reaction.

作者信息

Brito C M M, Lima M M, Sarquis O, Pires M Q, Coutinho C F S, Duarte R, Pacheco R S

机构信息

Departamento de Ciências Biológicas, Fundação Oswaldo Cruz, Escola Nacional de Saúde Pública, Rua Leopoldo Bulhões 1480, CP 926, Rio de Janeiro, 21041-210, Brazil.

出版信息

Parasitol Res. 2008 Oct;103(5):1111-7. doi: 10.1007/s00436-008-1102-5. Epub 2008 Jul 17.

Abstract

Different molecular markers have been employed for typing Trypanosoma cruzi strains from endemic areas of Chagas disease. The low-stringency single specific primer-polymerase chain reaction (LSSP-PCR) has been a sensitive and informative technique that uses the variable region of kinetoplast DNA minicircles as a genetic marker, allowing detection of DNA sequence variation. In the present study, we analyzed the intra-lineage genetic variability of the T. cruzi strains obtained from triatomine feces collected on filter paper FTA card by LSSP-PCR. The hybridization of the PCR products with a probe for the subgenus Schizotrypanum and a clone-specific probe from Dm28c confirmed the subgenus as T. (S.) cruzi and respective lineages as T. cruzi I. Phenetic analysis showed the presence of three clusters that diverged by different coefficients of similarity. Thirteen T. cruzi I genotypes were observed circulating among Triatoma pseudomaculata and Rhodnius nasutus from peridomiciliary and natural environments in five peri-urban and urban localities of Jaguaruana, Ceará, Brazil. These data indicate the importance of the circulation of T. cruzi I genotypes among T. pseudomaculata and R. nasutus in different environments and the possible risk of Chagas disease domestic transmission.

摘要

不同的分子标记已被用于对来自恰加斯病流行地区的克氏锥虫菌株进行分型。低严谨度单特异性引物聚合酶链反应(LSSP-PCR)是一种灵敏且信息丰富的技术,它利用动质体DNA微小环的可变区作为遗传标记,能够检测DNA序列变异。在本研究中,我们通过LSSP-PCR分析了从滤纸片FTA卡上收集的锥蝽粪便中获得的克氏锥虫菌株的谱系内遗传变异性。PCR产物与裂殖锥虫亚属探针以及来自Dm28c的克隆特异性探针杂交,证实该亚属为克氏锥虫(裂殖锥虫亚属),各谱系为克氏锥虫I。表型分析显示存在三个由不同相似系数分化而来的簇。在巴西塞阿拉州雅瓜拉纳的五个城郊和城市地区的住宅周围和自然环境中的伪红带锥蝽和鼻锥蝽中,观察到13种克氏锥虫I基因型在传播。这些数据表明克氏锥虫I基因型在不同环境中的伪红带锥蝽和鼻锥蝽之间传播的重要性以及恰加斯病家庭传播的潜在风险。

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