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

被引：1

作者：

Bhaskara, Adpangaya Sahana ^{[1
]}

Ahmed, Bandalli N. Jawad ^{[2
]}

Adkar-Purushothama, Charith R. ^{[3
]}

Prasad, Mysore N. Nagendra ^{[3
]}

Sreenivasa, Marikunte Y. ^{[1
]}

Maheshwar, Pavagada K. Murthy ^{[2
]}

机构：

[1] Univ Mysore, Dept Studies Microbiol, Mysore 570006, Karnataka, India

[2] Univ Mysore, Dept Microbiol, Yuvarajas Coll, Mysore 570005, Karnataka, India

[3] Sri Jayachamarajendra Coll Engn, Dept Biotechnol, Mysore 570 OO6, Karnataka, India

来源：

JOURNAL OF PLANT DISEASES AND PROTECTION | 2013年 / 120卷 / 5-6期

关键词：

hnPCR; Huanglongbing; Liberibacter; ribosomal protein gene; REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; GREENING DISEASE; ASIATICUS; AMPLIFICATION; IDENTIFICATION; BACTERIUM; GENES;

D O I：

10.1007/BF03356473

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

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 beta-operon (rp1KAJL-rpoBC) ribosomal protein genes from all the known 'Candidatus 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.

引用

页码：189 / 193

页数：5

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