RESUMO
Over the last decade, cancer genome sequencing has revealed in detail the genomic landscapes of an increasing number of common solid human tumors. This has greatly impacted the clinical care of cancer patients based on the fact that many of these tumors exhibit activating mutations in driver genes that are prone to target therapy, largely impacting cancer management strategies. Genomic heterogeneity of tumors is becoming an increasingly recognized phenomenon relevant to genome-based medicine. Because a large number tumors may display several mutations at the same time, multiplexing has become the preferred approach to reveal the mutational landscape in patient samples and to better design a targeted approach to their illness.
Assuntos
Técnicas de Genotipagem , Mutação , Neoplasias/genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único , Humanos , Espectrometria de Massas , Taxa de MutaçãoRESUMO
American Trypanosomiasis (Chagas disease) is an infectious disease caused by the hemoflagellate parasite Trypanosoma cruzi which is transmitted by reduviid bugs. T. cruzi infection occurs in a broad spectrum of reservoir animals throughout North, Central, and South America and usually evolves into an asymptomatic chronic clinical stage of the disease in which diagnosis is often challenging. This chapter describes the application of polymerase chain reaction (PCR) for the detection of Trypanosoma cruzi DNA including protocols for sample preparation, DNA extraction, and target amplification methods.
Assuntos
Doença de Chagas/diagnóstico , Doença de Chagas/parasitologia , Reação em Cadeia da Polimerase/métodos , Trypanosoma cruzi/genética , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Genes de Protozoários , Genes de RNAr , Humanos , Técnicas de Amplificação de Ácido NucleicoRESUMO
Leishmaniasis is an infectious disease caused by protozoan parasites of the genus Leishmania which are transmitted to humans through bites of infected sand flies. The variable clinical manifestations and the evolution of the disease are determined by the infecting species. Recognition at a species level is of utmost importance since this greatly impacts therapy decision making as well as predicts outcome for the disease. This chapter describes the application of polymerase chain reaction (PCR) in the detection of Leishmania parasites across the disease spectrum, including protocols for sample collection and transportation, genomic material extraction, and target amplification methods with special emphasis on PCR amplification of the cytochrome b gene for Leishmania spp. species identification.