<abstract xmlns="http://www.w3.org/1999/xhtml"><p> Phytosanitary measures to control diseases require sensitive and reliable detection methods. In case of Erwinia amylovora, the causal agent of fire blight, there are several methods developed. We tested six of them: (1) a conventional method; (2) standard PCR; (3) BIO-PCR; (4) Real-time PCR with SYBR Green; (5) Real-time PCR with TaqMan probe; and (6) Loop Mediated Isothermal Amplification (LAMP) in terms of their specificity, sensitivity and repeatability. Of the methods tested, only Real-time PCR with SYBR Green amplified non-specific fragments, which could be however identified by melting curve analysis. Real-time PCRs (both with SYBR Green and TaqMan probe) and LAMP were most sensitive and allowed to detect E. amylovora even in the samples where the concentration of cells was lower than 2 x 10<sup>1</sup> cfu/reaction. The highest repeatability was obtained using conventional method and Real-time PCRs. Of the methods used, only the LAMP technique was very insensitive to plant impurities. </p></abstract>

Phytosanitary measures to control diseases require sensitive and reliable detection methods. In case of Erwinia amylovora, the causal agent of fire blight, there are several methods developed. We tested six of them: (1) a conventional method; (2) standard PCR; (3) BIO-PCR; (4) Real-time PCR with SYBR Green; (5) Real-time PCR with TaqMan probe; and (6) Loop Mediated Isothermal Amplification (LAMP) in terms of their specificity, sensitivity and repeatability. Of the methods tested, only Real-time PCR with SYBR Green amplified non-specific fragments, which could be however identified by melting curve analysis. Real-time PCRs (both with SYBR Green and TaqMan probe) and LAMP were most sensitive and allowed to detect E. amylovora even in the samples where the concentration of cells was lower than 2 x 101 cfu/reaction. The highest repeatability was obtained using conventional method and Real-time PCRs. Of the methods used, only the LAMP technique was very insensitive to plant impurities.

Evaluation of methods for erwinia amylovora detection

Research Institute of Horticulture Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland

Journal of Horticultural Research

{"article-title":"Evaluation of methods for erwinia amylovora detection"}

Phytosanitary measures to control diseases require sensitive and reliable detection methods. In case of Erwinia amylovora, the causal agent of fire blight,...

Evaluation of methods for erwinia amylovora detection

Pubblicato online: 27 feb 2014

Pagine: 65 - 71

DOI: https://doi.org/10.2478/johr-2013-0023

Parole chiave
asymptomatic plant material, fire blight, LAMP, PCR, Real-time PCR

This content is open access.

Evaluation of methods for erwinia amylovora detection

Pubblicato online: 27 feb 2014

Pagine: 65 - 71

DOI: https://doi.org/10.2478/johr-2013-0023

Parole chiaveasymptomatic plant material, fire blight, LAMP, PCR, Real-time PCR

This content is open access.

Parole chiave
asymptomatic plant material, fire blight, LAMP, PCR, Real-time PCR