Highly sensitive detection and quantification of the pathogen Yersinia ruckeri in fish tissues by using real-time PCR

Bastardo, Asmine; Ravelo, Carmen; Romalde, Jesús L.

doi:10.1007/s00253-012-4328-1

Cited by 15 publications

(9 citation statements)

References 36 publications

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“…Improvements in specificity of both conventional and quantitative PCR (qPCR) assay have been attained using other candidate genes [ 39 - 41 ]. Bastardo et al [ 42 ] recently described a qPCR method based on the recombinant protein A (recA) gene . Gold nanoparticle-based assays are an emerging technology, which may become a powerful tool for rapid, direct and sensitive detection of unamplified Y. ruckeri nucleic acids in clinical samples.…”

Section: Diagnosismentioning

confidence: 99%

Yersinia ruckeri, the causative agent of enteric redmouth disease in fish

Kumar

Menanteau–Ledouble

Saleh

et al. 2015

Vet Res

157

136

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Enteric redmouth disease (ERM) is a serious septicemic bacterial disease of salmonid fish species. It is caused by Yersinia ruckeri, a Gram-negative rod-shaped enterobacterium. It has a wide host range, broad geographical distribution, and causes significant economic losses in the fish aquaculture industry. The disease gets its name from the subcutaneous hemorrhages, it can cause at the corners of the mouth and in gums and tongue. Other clinical signs include exophthalmia, darkening of the skin, splenomegaly and inflammation of the lower intestine with accumulation of thick yellow fluid. The bacterium enters the fish via the secondary gill lamellae and from there it spreads to the blood and internal organs. Y. ruckeri can be detected by conventional biochemical, serological and molecular methods. Its genome is 3.7 Mb with 3406–3530 coding sequences. Several important virulence factors of Y. ruckeri have been discovered, including haemolyin YhlA and metalloprotease Yrp1. Both non-specific and specific immune responses of fish during the course of Y. ruckeri infection have been well characterized. Several methods of vaccination have been developed for controlling both biotype 1 and biotype 2 Y. ruckeri strains in fish. This review summarizes the current state of knowledge regarding enteric redmouth disease and Y. ruckeri: diagnosis, genome, virulence factors, interaction with the host immune responses, and the development of vaccines against this pathogen.

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Section: Diagnosismentioning

confidence: 99%

Yersinia ruckeri, the causative agent of enteric redmouth disease in fish

Kumar

Menanteau–Ledouble

Saleh

et al. 2015

Vet Res

157

136

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“…Consequently, qPCR has been applied to the diagnosis of a wide range of pathogens from various sources including food products (Postollec, Falentin, Pavan, Combrisson & Sohier, 2011), faecal and environmental samples (B elanger, Boissinot, Clairoux, Picard & Bergeron, 2003;Rinttil€ a, Kassinen, Malinen, Krogius & Palva, 2004), and infected plant and animal tissues (Marancik & Wiens, 2013;Osman & Rowhani, 2006). These applications include assays for detection of Y. ruckeri (Argenton et al, 1996;Bastardo, Ravelo & Romalde, 2012), although commercial applicability of these particular methods and their value as a screening tool are limited, as they involve invasive or destructive sampling of putatively infected fish.…”

mentioning

confidence: 99%

A highly sensitive, non‐invasive qPCR‐based strategy for direct quantification of Yersinia ruckeri in fish faeces

Ghosh

Crosbie

Nowak

et al. 2018

Journal of Fish Diseases

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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.

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“…Tissue samples (kidney, liver and spleen), pooled from three fish for each outbreak, were aseptically collected for presumptive diagnosis of freshwater pathogens, including infectious pancreatic necrosis virus, Flavobacterium psychrophilum , and Y. ruckeri through PCR (Urdaci et al . ; Bastardo, Ravelo & Romalde ; Avendaño‐Herrera et al . ).…”

Section: Biochemical Characterization Of the Chilean Coho Salmon Isolmentioning

confidence: 99%

“…Therefore, amplifications were performed using previously described primer pairs (Versalovic, Koeuth & Lupski ) and protocols reported for Y. ruckeri (Bastardo et al . ), with the exception that PCRs were performed using the GoTaq Green Master Mix (Promega).…”

Section: Biochemical Characterization Of the Chilean Coho Salmon Isolmentioning

confidence: 99%

“…Enterobacterial repetitive intergenic consensus (ERIC-PCR) and repetitive extragenic palindromic (REP-PCR) can molecularly characterize bacterial pathogens in fish (Romalde 2005;Toranzo, Magariños & Romalde 2005). Therefore, amplifications were performed using previously described primer pairs (Versalovic, Koeuth & Lupski 1991) and protocols reported for Y. ruckeri (Bastardo et al 2012), with the exception that PCRs were performed using the GoTaq Green Master Mix (Promega). The three Chilean isolates returned identical fingerprinting profiles, regardless of the molecular tool used (i.e.…”

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confidence: 99%

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