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
收藏
页码：189 / 193
页数：5
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