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Evaluation of efficiency of hemi-nested PCR assay for the detection of ‘Candidatus Liberibacter’ infecting citrus

Abstract

The present study reports the development and evaluation of a hemi-nested polymerase chain reaction (hnPCR) assay for the efficient detection of ‘Candidatus Liberibacter’ infecting citrus plants. A set of new primers was designed by alignment of nucleotide sequences of the β-operon (rplKAJL-rpoBC) ribosomal protein genes from all the known ‘Candi-datus Liberibacter asiaticus’ isolates reported in Genbank. Hemi-nested PCR reaction components and thermal cycling parameters were optimized and reaction conditions were standardized. Sequencing of the PCR products from hemi nested-PCR reactions confirmed the specificity of new primer pairs for ‘Candidatus Liberibacter asiaticus’. The reliability and sensitivity of hnPCR was evaluated by comparing it to real-time PCR.

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Correspondence to Pavagada K. Murthy Maheshwar.

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Bhaskara, A.S., Ahmed, B.N.J., Adkar-Purushothama, C.R. et al. Evaluation of efficiency of hemi-nested PCR assay for the detection of ‘Candidatus Liberibacter’ infecting citrus. J Plant Dis Prot 120, 189–193 (2013). https://doi.org/10.1007/BF03356473

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Key words

  • hnPCR
  • Huanglongbing
  • Liberibacter
  • ribosomal protein gene