A simple method to generate PCR-RFLP typing profiles from DNA sequences in Toxoplasma gondii

Autor(es)

Pede Castro, Bruno Bello ^[1] ; Gennari, Solange Maria ^{[1, 2]} ; Lorenzi, Hernan ^[3] ; Su, Chunlei ^[4]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Prevent Vet Med & Anim Hlth, Fac Vet Med, Av Prof Orlando M de Paiva 87, BR-05408270 Sao Paulo, SP - Brazil

^[2] Univ Santo Amaro, Postgrad Program Vet Med, Av Eneas de Siqueira Neto 340, BR-04829300 Sao Paulo, SP - Brazil

^[3] J Craig Venter Inst, Rockville, MD 20895 - USA

^[4] Univ Tennessee, Dept Microbiol, Knoxville, TN 37996 - USA

• Tipo de documento: Artigo Científico
• Fonte: INFECTION GENETICS AND EVOLUTION; v. 85, NOV 2020.
• Citações Web of Science: 0

Resumo

Multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) has been widely used to genotype microorganisms. Accumulation of data generated by this method has helped to reveal genetic diversity, population structure and transmission of many microbial pathogens. Advances in DNA sequencing technologies have made it possible to identify microorganisms by multilocus sequencing typing (MLST) or whole genome sequence typing (WGST) to reach high resolution of identification. While MLST and WGST are gradually replacing PCR-RFLP for genotyping, invaluable databases generated by the latter may not be easily linked to datasets generated by sequencing based methods. In addition, DNA sequences corresponding to PCR-RFLP markers are often deposited in public domains but not fully explored to infer the RFLP profile. To alleviate this problem, we developed a simple protocol that can generate PCR-RFLP profiles from DNA sequence data, therefore facilitating the integration of data generated by different typing methods. Here we used the protozoan parasite Toxoplasma gondii as an example to bridge different typing methods. (AU)

• Processo FAPESP: 16/02646-3 - Desenvolvimento de uma nova geração de marcadores genéticos para estudos de epidemiologia molecular e genética de populações de Toxoplasma gondii
• Beneficiário: Bruno Bello Pede Castro
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado Direto