Liarte Daniel B, Murta Silvane M F, Steindel Mario, Romanha Alvaro J
Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisas René Rachou- Fiocruz, 30190-002 Belo Horizonte, Minas Gerais, Brazil.
Exp Parasitol. 2009 Dec;123(4):283-91. doi: 10.1016/j.exppara.2008.12.005. Epub 2008 Dec 16.
A multiplex PCR was developed for simultaneous detection of Trypanosoma cruzi DNA and classification of the parasite strain into groups I and II. As little as 10fg of T. cruzi DNA could be detected by multiplex PCR. The technique was shown to be specific for T. cruzi DNA, since no PCR amplification products were obtained with DNA from other tripanosomatid species. Multiplex PCR was validated by assaying genomic DNA from 34 strains of T. cruzi that had been previously characterized; 24 blood samples from experimentally-infected mice and non-infected controls; 20 buffy coat samples from patients in the acute phase of Chagas disease and non-infected individuals, and 15 samples of feces from naturally-infected Triatoma infestans. T. cruzi samples from patients and from Y strain-infected mice were classified by multiplex PCR as T. cruzi II and samples from T. infestans and Colombiana strain-infected mice as T. cruzi I.
开发了一种多重PCR方法，用于同时检测克氏锥虫DNA并将寄生虫菌株分为I组和II组。多重PCR可检测低至10fg的克氏锥虫DNA。该技术对克氏锥虫DNA具有特异性，因为来自其他锥虫物种的DNA未获得PCR扩增产物。通过检测34株先前已鉴定的克氏锥虫的基因组DNA、24份来自实验感染小鼠和未感染对照的血样、20份来自恰加斯病急性期患者和未感染个体的血沉棕黄层样本以及15份来自自然感染的骚扰锥蝽的粪便样本，对多重PCR进行了验证。通过多重PCR将来自患者和Y株感染小鼠的克氏锥虫样本分类为克氏锥虫II型，将来自骚扰锥蝽和哥伦比亚株感染小鼠的样本分类为克氏锥虫I型。
