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Trypanosoma cruzi: Multiplex PCR to detect and classify strains according to groups I and II |
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| Authors: | Daniel B Liarte Silvane MF Murta Alvaro J Romanha |
| Institution: | a Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisas René Rachou- Fiocruz, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, Minas Gerais, Brazil b Departamento de Microbiologia e Parasitologia, Universidade Federal de Santa Catarina, 88040-900 Florianópolis, Santa Catarina, Brazil |
| Abstract: | 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 10 fg 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. |
| Keywords: | Trypanosoma cruzi Satellite DNA Multiplex PCR Molecular diagnosis T cruzi group Triatomine Triatoma infestans rDNA ribosomal DNA DNA deoxyribonucleic acid kDNA kinetoplast deoxyribonucleic acid PBS phosphate buffered saline PCR polymerase chain reaction RAPD random amplification of polymorphic DNA RFLP restriction fragment length polymorphism T cruzi I Trypanosoma cruzi group I T cruzi II Trypanosoma cruzi group II Z1 Z2 Z3 zymodemes 1 2 and 3 |
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