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

被引：15

作者：

Adkar-Purushothama, Charith Raj ^{[1
]}

Maheshwar, P. K. ^{[2
]}

Sano, Teruo ^{[3
]}

Janardhana, G. R. ^{[1
]}

机构：

[1] Univ Mysore, Dept Studies Bot, Mol Phytodiagnost Lab, Mysore 570006, Karnataka, India

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

[3] Hirosaki Univ, Fac Agr & Life Sci, Plant Pathol Lab, Hirosaki, Aomori, Japan

来源：

CURRENT MICROBIOLOGY | 2011年 / 62卷 / 05期

关键词：

REAL-TIME PCR; REVERSE TRANSCRIPTION; DIFFERENTIATION; STRAINS; ISOLATE; APHIDS;

D O I：

10.1007/s00284-011-9883-7

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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.

引用

页码：1455 / 1459

页数：5

共 24 条

[1] Characterization of the mixture of genotypes of a Citrus tristeza virus isolate by reverse transcription-quantitative real-time PCR [J].

Ananthakrishnan, G. ;

Venkataprasanna, T. ;

Roy, Avijit ;

Brlansky, R. N. .

JOURNAL OF VIROLOGICAL METHODS, 2010, 164 (1-2) :75-82

[2] THE CONTINUOUS CHALLENGE OF CITRUS TRISTEZA VIRUS CONTROL [J].

BARJOSEPH, M ;

MARCUS, R ;

LEE, RF .

ANNUAL REVIEW OF PHYTOPATHOLOGY, 1989, 27 :291-316

[3] INSITU IMMUNOFLUORESCENCE FOR THE DETECTION OF CITRUS TRISTEZA VIRUS INCLUSION-BODIES [J].

BRLANSKY, RH ;

LEE, RF ;

GARNSEY, SM .

PLANT DISEASE, 1988, 72 (12) :1039-1041

[4]

Cambra M., 2000, Proceedings of the 14th Conference of the International Organization of Citrus Virologists, Campinas, Sao Paulo State, Brazil, 13-18 September 1998, P42

[5] Incidence and epidemiology of Citrus tristeza virus in the Valencian Community of Spain [J].