采用 DNA 探针杂交和特异性聚合酶链反应（PCR）对来自阿根廷东北部地区人类样本中的克氏锥虫亚谱系进行基因分型。
Genotyping of Trypanosoma cruzi sublineage in human samples from a North-East Argentina area by hybridization with DNA probes and specific polymerase chain reaction (PCR).
机构信息
Laboratorio de Tecnología Inmunológica, Facultad de Bioquímica y Cs Biológicas, Universidad Nacional del Litoral, Argentina.
出版信息
Am J Trop Med Hyg. 2010 Jan;82(1):67-73. doi: 10.4269/ajtmh.2010.09-0391.
Abstract
We have evaluated blood samples of chronic and congenital Trypanosoma cruzi-infected patients from the city of Reconquista located in the northeast of Argentina where no information was previously obtained about the genotype of infecting parasites. Fourteen samples of congenital and 19 chronical patients were analyzed by hybridization with DNA probes of minicircle hypervariable regions (mHVR). In congenital patients, 50% had single infections with TcIId, 7% single infections with TcIIe, 29% mixed infections with TcIId/e, and 7% had mixed infections with TcIId/b and 7% TcIId/b, respectively. In Chronical patients, 52% had single infections with TcIId, 11% single infections with TcIIe, 26% had mixed infections with TcIId/e, and 11% had non-identified genotypes. With these samples, we evaluated the minicircle lineage-specific polymerase chain reaction assay (MLS-PCR), which involves a nested PCR to HVR minicircle sequences and we found a correlation with hybridization probes of 96.4% for TcIId and 54.8% for TcIIe.
摘要
我们评估了来自阿根廷东北部雷孔基斯塔市的慢性和先天性克氏锥虫感染患者的血液样本，此前该地区尚无关于感染寄生虫基因型的信息。通过与微环高变区（mHVR）的 DNA 探针杂交，分析了 14 例先天性和 19 例慢性患者的样本。在先天性患者中，50%为 TcIId 单一感染，7%为 TcIIe 单一感染，29%为 TcIId/e 混合感染，7%为 TcIId/b 和 7%的 TcIId/b 混合感染。在慢性患者中，52%为 TcIId 单一感染，11%为 TcIIe 单一感染，26%为 TcIId/e 混合感染，11%为未鉴定基因型。用这些样本，我们评估了微环谱系特异性聚合酶链反应检测（MLS-PCR），该检测涉及到对 HVR 微环序列的巢式 PCR，我们发现与 TcIId 的杂交探针的相关性为 96.4%，与 TcIIe 的相关性为 54.8%。
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