Sensitive and Reliable RT-Nested PCR Assay for Detection of Citrus tristeza Virus from Naturally Infected Citrus Plants

A specific and sensitive reverse transcriptase-nested polymerase chain reaction assay (RT-nPCR) was developed for the detection of Citrus tristeza virus (CTV) from naturally infected citrus samples. Two sets of primer pairs were designed by alignment of nucleotide sequences available in GenBank data...

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Bibliographic Details
Published in:Current microbiology 2011-05, Vol.62 (5), p.1455-1459
Main Authors: Adkar-Purushothama, Charith Raj, Maheshwar, P. K, Sano, Teruo, Janardhana, G. R
Format: Article
Language:English
Subjects:
Bioassays
Biomedical and Life Sciences
Biotechnology
Citrus
Citrus - virology
Citrus fruits
Citrus tristeza virus
Closterovirus - genetics
Closterovirus - isolation & purification
DNA Primers - genetics
Fruits
genotype
Genotypes
Life Sciences
Microbiology
monitoring
nucleotide sequences
orchards
Plant diseases
Plant Diseases - virology
Plant pathology
Polymerase chain reaction
Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction - methods
Virology
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Summary:A specific and sensitive reverse transcriptase-nested polymerase chain reaction assay (RT-nPCR) was developed for the detection of Citrus tristeza virus (CTV) from naturally infected citrus samples. Two sets of primer pairs were designed by alignment of nucleotide sequences available in GenBank database for different genotypes of CTV. RT-nPCR reaction components and thermal cycling parameters were optimized and reaction conditions were standardized. Sequencing of the PCR products from direct and nested-PCR reactions confirmed the specificity of both primer pairs. Presence of CTV specific amplicons in asymptomatic samples which were collected from diseased orchards indicated the sensitivity of the test. As RT-nPCR technique, developed in the present study, is specific and efficient in detecting CTV, this could be envisioned for diagnostic applications and surveillance.
ISSN:0343-8651
1432-0991
DOI:10.1007/s00284-011-9883-7