An RT-PCR procedure for detection and surveillance of Citrus leprosis virus C (CiLV-C) in post-entry quarantine stocks of citrus.

• Autor(a) principal: OCHOA-CORONA, F. M.
• Data de Publicação: 2009
• Outros Autores: LOCALI-FABRIS, E. C., FREITAS-ASTÚA, J.
• Idioma: eng
• Título da fonte: Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
• Texto Completo: http://www.alice.cnptia.embrapa.br/alice/handle/doc/748515
• topic: Brevipalpus Phoenicis; Fruta Cítrica; Citrus

Citrus leprosis virus C (CiLV-C), which is transmitted both mechanically and by the mite Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae), causes significant disease damage in South and Central America. Citrus leprosis disease was first recorded in Florida in 1925, but is believed eradicated through improved mite control procedures. CiLV-C is a threat to citrus producing nations where it is not present, such as New Zealand and the USA, and a sensitive detection method is required for screening and biosecurity of suspect quarantine material. CiLV-C is mechanically transmitted, posses a bipartite RNA genome and was believed to be a rhabdovirus. After being sequenced, CiLV-C was proposed as the type member of a new genus, Cilevirus, related to several (+) ssRNA viruses. Of two known morphological types of CiLV particles, the cytoplasmic type (CiLV-C) is prevalent than the nuclear type (CiLV-N) in Brazil and elsewhere. A pair of diagnostic primers, amplifying a segment of 278 bp located at the RNA-2 p15 gene of CiLV-C was designed using the Web software pathway Primer3-mFOLD-BLASTn. A thermodynamically robust RT-PCR that performs well in a range of melting temperatures and specifically optimized for CiLV-C was developed, and is a feasible tool to be used in quarantine.