Print-capture PCR for detection of tomato begomoviruses from plants and whiteflies

• Autor(a) principal: Nagata,Tatsuya
• Data de Publicação: 2004
• Outros Autores: Inoue-Nagata,Alice K, Ávila,Antonio C. de, Giordano,Leonardo de B
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Fitopatologia Brasileira
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582004000100014
• topic: Geminivírus; Bemisia tabaci; Bemisia argentifoli

Print-capture (PC) Polymerase chain reaction (PCR) was evaluated as a novel detection method of plant viruses. Tomato (Lycopersicon esculentum) plants infected with begomovirus (fam. Geminiviridae, gen. Begomovirus) and viruliferous whiteflies were used to study the efficiency of the method. Print-capturing steps were carried out using non-charged nylon membrane or filter paper as the solid support for DNA printings. Amplified DNA fragments of expected size were consistently obtained by PCR from infected plants grown in a greenhouse, after direct application of printed materials to the PCR mix. However, virus detection from a single whitefly and from field-grown tomato samples required a high temperature treatment of printed material prior to PCR amplification. Comparison of nylon membrane and filter paper as the solid support revealed the higher efficiency of the nylon membrane. The application of print-capture PCR reduces the chances of false-positive amplification by reducing manipulation steps during preparation of the target DNA. This method maintains all the advantages of PCR diagnosis, such as the high sensitivity and no requirement of radioactive reagents.