Immune Tissue Print and Immune Capture-PCR for Diagnosis and Detection of Candidatus Liberibacter Asiaticus

Sci Rep. 2017 Apr 18:7:46467. doi: 10.1038/srep46467.

Abstract

'Candidatus Liberibacter asiaticus' (CaLas), associated with citrus Huanglongbing (HLB), is a non culturable member of the α-proteobacteria. In this study serologically based methods for the detection of CaLas were developed. An anti-outer membrane protein A (OmpA) polyclonal antibody previously produced (in our laboratory) was highly effective for the detection of CaLas from citrus tissues in a simple tissue printing format. The antibody was also used to capture bacteria from periwinkle extracts. About 80% of all field samples analyzed tested positive with both immune tissue printing and qPCR; whereas 95% were positive with at least one of these two methods. When asymptomatic citrus tissues were tested, the tissue printing method gave a higher rate of detection (83%) than the qPCR method (64%). This is consistent with a lower concentration of CaLas DNA, but a higher proportion of viable cells, in the asymptomatic tissues. The immune tissue printing method also highlights the detail of the spatial distribution of 'Ca. Liberibacter asiaticus' in diseased citrus tissues. Both the immune capture PCR and immune tissue printing methods offer the advantages of low cost, high throughput, ease of scaling for multiple samples and simplicity over current PCR-based methods for the detection of 'Ca. Liberibacter asiaticus'.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Bacterial / metabolism*
  • Bacterial Outer Membrane Proteins / immunology*
  • Citrus / immunology
  • Citrus / microbiology*
  • DNA, Bacterial / analysis
  • Plant Diseases / microbiology*
  • Polymerase Chain Reaction
  • Rhizobiaceae / genetics
  • Rhizobiaceae / immunology*
  • Sensitivity and Specificity
  • Tissue Distribution

Substances

  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • DNA, Bacterial
  • OMPA outer membrane proteins