Detection of Haemophilus parasuis isolates from South China by loop-mediated isothermal amplification and isolate characterisation.
Onderstepoort J Vet Res
; 79(1): E1-6, 2012 Apr 24.
Article
em En
| MEDLINE
| ID: mdl-23327310
ABSTRACT
Haemophilus parasuis is the etiological agent of Glässer's disease, which is characterised by ï¬brinous polyserositis, meningitis and polyarthritis, causing severe economic losses to the swine industry. In this study, a loop-mediated isothermal ampliï¬cation (LAMP) test was developed to improve the speciï¬city, facility and speed of diagnosis of H. parasuis isolates. The LAMP assay rapidly ampliï¬ed the target gene within 50 min incubation at 63 °C in a laboratory water bath. The LAMP amplicon could be visualised directly in the reaction tubes following the addition of SYBR Green I dye. The detection limit of this LAMP method was 10 CFU/mL, which was 10 times more sensitive than the earlier 16S rRNA polymerase chain reaction (PCR) test conducted by Oliveira, Galina and Pijoan (2001), and no cross-reactivity was observed from other non-H. parasuis strains. This LAMP test was evaluated further on 187 clinical specimens from pigs suspected of being infected with H. parasuis. Forty-three were found positive by bacterial isolation of H. parasuis, as well as by the 16S rRNA PCR and LAMP tests. The 43 H. parasuis isolates were classiï¬ed into 9 serovars and had 37 genetic patterns when analysed by pulsed-ï¬eld gel electrophoresis (PFGE). This displayed that various H. parasuis serovars and genotypes were widely distributed in South China. Therefore, the speed, speciï¬city and sensitivity of the LAMP test, the lack of a need for expensive equipment, and the visual readout showed great potential for a correct clinical diagnosis of H. parasuis in favour of controlling Glässer's disease.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Doenças dos Suínos
/
Técnicas de Amplificação de Ácido Nucleico
/
Haemophilus parasuis
/
Infecções por Haemophilus
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
País como assunto:
Asia
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article