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

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Author(s): OCHOA-CORONA, F. M.; LOCALI-FABRIS, E. C.; FREITAS-ASTÚA, J.

Summary: 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.

Publication year: 2009

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