Comparison of specificity and sensitivity of immunochemical and molecular techniques for reliable detection of Erwinia amylovora.
Folia Microbiol (Praha)
; 52(2): 175-82, 2007.
Article
en En
| MEDLINE
| ID: mdl-17575916
Erwinia amylovora [(BURRILL) WINSLOW et al.] (Ea), the causal agent of fire blight, was detected in plant samples and pure bacterial cultures by means of PCR, IFAS and ELISA. Polyclonal antibodies of Neogen Europe Ltd. were used for IFAS and PTA-ELISA and laboratory-generated primers EaF72 and EaR560 for PCR. Using the BIOLOG system and an immature pear fruit assay, identities of all Ea strains were confirmed as the fire blight bacterium. In assays of pure Ea cultures, PTA-ELISA, and both IFAS and PCR were sensitive to concentrations 10(6)-10(5) and 10(5)-10(4) CFU/mL, respectively. When saprophytic bacteria associated with Ea in plant samples were tested as potentially cross-reacting bacteria, PTA-ELISA and IFAS gave 20 and 14 % cross-reactions, respectively. In plant samples, the presence of Ea was more reliably detected by IFAS (at a dilution of 1 : 1000) than by PTA-ELISA (to dilution 1 : 100). The capacity to detect Ea might be increased using an optimized PCR, but for PCR prepared from infected plant samples it was necessary to use the bacterial DNA isolated with a DNeasy Plant Mini Kit (Qiagen). In this case the PCR was sensitive to a concentration of 10(5) CFU/mL. PCR was much more specific than either immunochemical technique, because no false positives were observed when primers EaF72 and EaR560 were used.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Enfermedades de las Plantas
/
Ensayo de Inmunoadsorción Enzimática
/
Reacción en Cadena de la Polimerasa
/
Técnica del Anticuerpo Fluorescente Indirecta
/
Erwinia amylovora
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Folia Microbiol (Praha)
Año:
2007
Tipo del documento:
Article
Pais de publicación:
Estados Unidos