A highly sensitive, non-invasive qPCR-based strategy for direct quantification of Yersinia ruckeri in fish faeces.
J Fish Dis
; 41(9): 1421-1428, 2018 Sep.
Article
en En
| MEDLINE
| ID: mdl-29926930
ABSTRACT
Finfish with asymptomatic Yersinia ruckeri infections pose a major risk as they can transmit the pathogen and cause clinical outbreaks in stock populations. Current tools have insufficient quantitative ability for accurately detecting the trace levels of Y. ruckeri typically associated with asymptomatic infection, necessitate invasive or lethal sampling, or require long processing times. This study presents a highly sensitive qPCR-based method, targeting part of the Y. ruckeri 16S rRNA sequence, that is capable of detecting extremely low levels of Y. ruckeri in noninvasively collected faecal samples. Quantitative precision and accuracy of faecal sample analysis was consistent, despite the complexity of the faecal matrix. The assay demonstrated linearity over a six log-wide dynamic range. Its limit of detection (LOD) and limit of quantification (LOQ) were 4 and 10 copies of the target sequence, respectively. Sensitivity of the assay was comparable to other qPCR-based methods without requiring invasive or lethal sampling. Applicability as a screening strategy was tested using passively collected faecal samples. Asymptomatic Y. ruckeri infection was detected in all samples, although none of the fish exhibited overt infection. This method will be beneficial for finfish disease management if developed further as a noninvasive, screening tool against asymptomatic Y. ruckeri infection.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Yersiniosis
/
Oncorhynchus mykiss
/
Yersinia ruckeri
/
Heces
/
Reacción en Cadena en Tiempo Real de la Polimerasa
/
Enfermedades de los Peces
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
J Fish Dis
Asunto de la revista:
BIOLOGIA
/
MEDICINA VETERINARIA
Año:
2018
Tipo del documento:
Article
País de afiliación:
Australia