Comparison of Traditional and Molecular Methods for Detection of Renibacterium Salmoninarum

Pascho, Ronald J.; Elliott, Diane G.; Chase, Dorothy M.

doi:10.1007/978-94-017-2315-2_7

Cited by 22 publications

(43 citation statements)

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“…21 The ELISA has been reported to be more sensitive than bacteriologic culture or the fluorescent antibody test (FAT) for detection of R. salmoninarum in fish tissues. 17 Although the ELISA has become a standard method recommended for large-scale screening of fish populations for R. salmoninarum, 13,16 certain problems can cause inconsistent results. For example, the quality of antibody lots used for this assay can be variable, 22 R. salmoninarum antigen can persist in fish in the absence of live bacteria, 20 and cross-reactivity with other bacterial species has been reported for some ELISA procedures.…”

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“…17 For confirmation of ELISA results and for primary diagnosis, a number of polymerase chain reaction (PCR) methods for R. salmoninarum detection have been developed. 17 Most of these PCR methods amplify segments of the msa gene from genomic DNA, although procedures to detect other nucleic acid sequences have been described. 17 Further improvements in PCR sensitivity have been obtained by a nested amplification of specific DNA sequences.…”

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confidence: 99%

“…17 Most of these PCR methods amplify segments of the msa gene from genomic DNA, although procedures to detect other nucleic acid sequences have been described. 17 Further improvements in PCR sensitivity have been obtained by a nested amplification of specific DNA sequences. 17 Direct comparisons of samples from naturally infected fish by use of ELISA, FAT, and PCR analysis have indicated higher sensitivity of a nested PCR than that of both of the other tests for detection of R. salmoninarum in kidney tissue 4 and coelomic fluid.…”

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“…17 Further improvements in PCR sensitivity have been obtained by a nested amplification of specific DNA sequences. 17 Direct comparisons of samples from naturally infected fish by use of ELISA, FAT, and PCR analysis have indicated higher sensitivity of a nested PCR than that of both of the other tests for detection of R. salmoninarum in kidney tissue 4 and coelomic fluid. 15 Although PCR procedures may be more sensitive than ELISA for R. salmoninarum detection, they differ from ELISA in that conventional PCR analyses cannot quantify infection levels and, therefore, cannot be used to determine the severity of infection in R. salmoninarum-positive fish in a population, or to monitor changes in infection levels over time or in response to various treatments.…”

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“…These bacteria included obligate and opportunistic fish pathogens, species reported to cross-react with R. salmoninarum in immunodiagnostic tests, and species phylogenetically similar to R. salmoninarum. 17 The bacteria were cultured as described. 4 Kidney tissue was obtained from adult male and female fall Chinook salmon collected from the Root River, Wisconsin weir during the fall of 2002 and tested for R. salmoninarum by nested PCR analysis and ELISA to obtain R. salmoninarum-negative tissue.…”

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Detection and Quantification of Renibacterium Salmoninarum DNA in Salmonid Tissues by Real-Time Quantitative Polymerase Chain Reaction Analysis

2006

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Abstract. Renibacterium salmoninarum is an important salmonid pathogen that is difficult to culture. We developed and assessed a real-time, quantitative, polymerase chain reaction (qPCR) assay for the detection and enumeration of R. salmoninarum. The qPCR is based on TaqMan technology and amplifies a 69-base pair (bp) region of the gene encoding the major soluble antigen (MSA) of R. salmoninarum. The qPCR assay consistently detected as few as 5 R. salmoninarum cells per reaction in kidney tissue. The specificity of the qPCR was confirmed by testing the DNA extracts from a panel of microorganisms that were either common fish pathogens or reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA). Kidney samples from 38 juvenile Chinook salmon (Oncorhynchus tshawytscha) in a naturally infected population were examined by real-time qPCR, a nested PCR, and ELISA, and prevalences of R. salmoninarum detected were 71, 66, and 71%, respectively. The qPCR should be a valuable tool for evaluating the R. salmoninarum infection status of salmonids.

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Detection and Quantification of Renibacterium Salmoninarum DNA in Salmonid Tissues by Real-Time Quantitative Polymerase Chain Reaction Analysis

2006

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Vaccination against Bacterial Kidney Disease

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Comparison of injection and immersion challenges of Renibacterium salmoninarum strains in Rainbow Trout

Watson

Bruce

et al. 2023

J Aqua Anim Hlth

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Objective Renibacterium salmoninarum is a pathogenic gram‐positive bacterium and is the causative agent of bacterial kidney disease (BKD), a malady that mainly impacts salmonid species. Experimental challenges were conducted to assess the virulence and challenge route for select R. salmoninarum strains (CK‐90 and ATCC 33739) in Rainbow Trout Oncorhynchus mykiss. Methods The CK‐90 strain was intracoelomically injected (100 μL) at a high dose containing 4.80 × 106 CFU/g of fish (optical density at 525 nm [OD525] = 1.779) and a low dose containing 6.86 × 105 CFU/g of fish (OD525 = 1.077); alternatively, fish were immersed in a solution containing 4.5 × 107 CFU/mL of fish (OD525 = 0.886). The ATCC 33739 strain (originating from Brook Trout Salvelinus fontinalis) was also included and intracoelomically injected at 3.58 × 105 CFU/g of fish (OD525 = 1.431) to discern differences in virulence between the strains. Result Clinical signs of BKD manifested at approximately 10 d postchallenge, and mortalities began at 19 days postchallenge. To confirm infection and quantify R. salmoninarum antigen load, an enzyme‐linked immunosorbent assay (ELISA) was conducted using kidney tissue collected after the challenge. Rainbow Trout that were challenged with CK‐90 by injection (both high‐ and low‐dose groups) exhibited significantly higher mortality than fish that were injected with ATCC 33739 or those that were exposed to CK‐90 via immersion challenge. The R. salmoninarum p57 (57‐kDa protein) antigen was confirmed via ELISA. Antigen load for fish injected with CK‐90 (high dose: OD405 = 0.71; low dose: OD405 = 0.66) was significantly higher than that for fish injected with ATCC 33739 (OD405 = 0.34). The CK‐90 strain (both high and low doses) was more virulent than ATCC 33739, which caused no mortalities over the 28‐days trial. Although there were no mortalities among ATCC 33739 fish, the ELISA confirmed that the R. salmoninarum antigen infiltrated kidney tissue in those fish. Conclusion The immersion challenge methodology for R. salmoninarum CK‐90 was ineffective for inducing mortalities at the examined dose.

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Comparison of Traditional and Molecular Methods for Detection of Renibacterium Salmoninarum

Cited by 22 publications

References 210 publications

Detection and Quantification of Renibacterium Salmoninarum DNA in Salmonid Tissues by Real-Time Quantitative Polymerase Chain Reaction Analysis

Detection and Quantification of Renibacterium Salmoninarum DNA in Salmonid Tissues by Real-Time Quantitative Polymerase Chain Reaction Analysis

Vaccination against Bacterial Kidney Disease

Comparison of injection and immersion challenges of Renibacterium salmoninarum strains in Rainbow Trout

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