Citrus psorosis virus: nucleotide sequencing of the coat protein gene and detection by hybridization and RT-PCR.

Abstract

Citrus psorosis virus (CPV) is a multicomponent

Recent advances in Citrus psorosis virus

“…RNAs 1, 2 and 3 of the isolate CPV-4 have been sequenced [5,39,58,59]. RNA extracted from citrus infected tissue is assayed for the presence of CPsV sequences by RT-PCR, cDNA copies are made and amplified for analyses.…”

“…The CPG has been used to develop RT-PCR [5]. In both cases, the RT-PCR was able to detect CPsV at concentrations substantially lower than those required for detection by immunoassays and electron microscopy [20].…”

“…Some isolates of CPsV that could be detected by RT-PCR could not be detected by hybridizations assays due to low titre of the virus. Nevertheless, these isolates were not detected at hybridization temperatures lower than 55°C due to non-specific reactions with host nucleic acids [5].…”

“…The tools used to detect CPsV in citrus trees are mainly based on using indicator plants and on laboratory tests including enzyme-linked immunosorbent assay (ELISA) [20,33,80]. After CPsV sequences became available, molecular tests based on RT-PCR and dot blot or tissue-print hybridization were also developed for CPsV detection [5,20,26,28,32,55,57,67].…”

“…CPsV can be detected by dot blot or tissue print hybridization using fulllength 32 P-labelled riboprobes of the CPG [5]. Positive sense probes hybridized to several isolates in tissue print and dot-blots at 55°C.…”