JCM Figure table search 04
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Moral, C. H.
Right arrow Articles by Carrasco, G. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Moral, C. H.
Right arrow Articles by Carrasco, G. N.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 1999, p. 1575-1578, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Molecular Cloning and Sequencing of the aroA Gene from Actinobacillus pleuropneumoniae and Its Use in a PCR Assay for Rapid Identification

Carmen Hernanz Moral, Alberto Cascón Soriano, María Sánchez Salazar, Javier Yugueros Marcos, Susana Suárez Ramos, and German Naharro Carrasco*

Departamento de Sanidad Animal, Microbiología e Inmunología, Facultad de Veterinaria, Universidad de León, 24071 León, Spain

Received 9 October 1998/Returned for modification 9 December 1998/Accepted 29 January 1999

The gene (aroA) of Actinobacillus pleuropneumoniae, serotype 2, encoding 5-enolpyruvylshikimate-3-phosphate synthase was cloned by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829, and the nucleotide sequence was determined. A pair of primers from the 5' and 3' termini were selected to be the basis for development of a specific PCR assay. A DNA fragment of 1,025 bp was amplified from lysed A. pleuropneumoniae serotypes 1 to 12 of biovar 1 or from isolated DNA. No PCR products were detected when chromosomal DNAs from other genera were used as target DNAs; however, a 1,025-bp DNA fragment was amplified when Actinobacillus equuli chromosomal DNA was used as a target, which could be easily differentiated by its NAD independence. The PCR assay developed was very sensitive, with lower detection limits of 12 CFU with A. pleuropneumoniae cells and 0.8 pg with extracted DNA. Specificity and sensitivity make this PCR assay a useful method for the rapid identification and diagnosis of A. pleuropneumoniae infections.

* Corresponding author. Mailing address: Departamento de Sanidad Animal, Microbiología e Inmunología, Facultad de Veterinaria, Universidad de Léon, 24071 Léon, Spain. Phone: 34 987 291 294. Fax: 34 987 291 304. E-mail: dsagnc{at}unileon.es.

Journal of Clinical Microbiology, May 1999, p. 1575-1578, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
Clin. Vaccine Immunol. ALL ASM JOURNALS

Copyright © 1999 by the American Society for Microbiology. All rights reserved.