Detection of<i>Renibacterium salmoninarum</i>Infection in Asymptomatic Atlantic Salmon

Lovely, Jane E.; Cabo, Carmen; Griffiths, Steven G.; Lynch, William H.

doi:10.1577/1548-8667(1994)006<0126:dorsii>2.3.co;2

Cited by 25 publications

(17 citation statements)

References 11 publications

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“…In ELISA and MF-FAT assays the quantitative correlation between culturable or viable cells and the measurement values is unclear. Culture on selective kidney disease medium (SKDM) (Austin et al 1983) is more relevant and sensitive for the detection of culturable cells (Olivier et al 1992, Lovely et al 1994, Griffiths et al 1996, but long incubation times (12 to 19 wk) are required to detect macroscopic colonies (Benediktsdóttir et al 1991). This makes the culture technique less practical for BKD diagnostic and screening purposes (Gudmundsdóttir et al 1993).…”

Section: Introductionmentioning

confidence: 99%

Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta

Suzuki

Sakai²

2007

Dis. Aquat. Org.

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Quantification of msa gene mRNA of Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD), was investigated using reverse transcription followed by real-time PCR assay on R. salmoninarum in culture, and in experimentally challenged chum salmon Oncorhynchus keta fry kidney tissues (total of 70 samples) after intraperitoneal (i.p.) injection and bath infection. Correlations of msa gene mRNA concentrations with culturable cell concentrations (as colony forming units [CFU]), determined by drop-plate culture method on selective kidney disease medium (SKDM) agar through a 12 wk incubation time, and msa gene DNA concentrations by realtime PCR assay were examined. Furthermore, ovarian fluid samples from wild chum salmon adults with no clinical signs of disease were collected from 8 rivers and from clinically infected kokanee O. nerka and masu salmon O. masou that were reared in 1 and 2 hatcheries, respectively (total of 414 samples). All samples were examined by nested PCR assay. Then, positive samples were examined by real-time PCR assays for mRNA and DNA; mRNA was detectable at 8 log units (5.0 × 10 1 to 5.0 × 10 9 copies µl -1 ) with high correlation (R 2 = 0.999). The mRNA concentration correlated with CFU in kidney tissue from fish infected by i.p. injection (R 2 = 0.924), by bath infection (R 2 = 0.502) and in culture (R 2 = 0.888). R. salmoninarum was detected and quantified by real-time PCR assay for mRNA in ovarian fluid samples in both subclinically infected chum salmon adults and clinically infected kokanee and masu salmon adults; detection rates ranged from 0 to 44.4% and concentrations ranged from 9.7 × 10 2 to 5.6 × 10 5 copies µl -1. These results indicate that real-time PCR assay for the mRNA is a rapid, sensitive and reliable method to detect and quantify the viability of R. salmoninarum in kidney and ovarian fluid samples of salmonid fishes with both clinical and subclinical infection of the pathogen.

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Section: Introductionmentioning

confidence: 99%

Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta

Suzuki

Sakai²

2007

Dis. Aquat. Org.

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“…; Lovely et al . ; Starliper & Teska ; Jónsdóttir et al . ), and evidence suggests that infected fish do not inevitably succumb to disease, but can eliminate the pathogen under certain conditions (Sanders, Pilcher & Fryer ; Bruno ; Lovely et al .…”

Section: Introductionmentioning

confidence: 99%

Bench‐top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture

Elliott

Applegate

Murray

et al. 2013

Journal of Fish Diseases

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No gold standard assay exhibiting error-free classification of results has been identified for detection of Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non-culture assays in three matrices (phosphate-buffered saline, ovarian fluid and kidney tissue). Non-culture assays included polyclonal enzyme-linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane-filtration FAT, nested polymerase chain reaction (nested PCR) and three real-time quantitative PCR assays. Injection challenge of specific pathogen-free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.

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“…challenge at a temperature of 12°C. However, this method was not found to be reliable for the identification of Rs-exposed salmon without clinical signs of BKD at temperatures below 8 O C (Lovely et al 1994). Sakai et al (1991) developed a polyclonal ELISA for the detection of serum antibodies to Rs in coho salmon Oncorhynchus kisutch.…”

Section: Introductionmentioning

confidence: 99%

Detection of humoral antibodies to Renibacterium salmoninarum in rainbow trout Oncorhynchus mykiss and Atlantic salmon Salmo salar challenged by immersion and in naturally infected populations

Jansson¹,

Ljungberg²

1998

Dis. Aquat. Org.

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Humoral antibodies to heat-stable antigens of Renibacterium salmoninar~~m (Rs) were detected by enzyme-linked immunosorbent assay (ELIS,I) in rainbow trout Oncorhynchus mykiss and in Atlantic salmon Salmo salar challenged by immersion A slow antibody response was found: 3%(1/30) was positive 4 wk after immersion and 7 2 % (26/36) was positive after 8 wk. All 30 fish sampled after 4 wk were found to b e infected, as determined by bacterial culture and/or the presence of soluble antigens in the kidney. At G, 8 and 12 wk after immersion the proportion of positives indicated by ELlSA was 58Ytp. The Rs infection was detected by cultivation in 3 6 % of sampled fish collected on the same occasion. Elevated antibody titres to Rs were detected in samples from both Atlantic salinon (59% In 1 farm) and from rainbow trout (20%) in 1 of 5 sampled farms) in naturally exposed populations all of which classified posltive for bacterial kidney disease (BKD). Elevated antibody titres were detected among sampled fish from populations of rainbow trout and salmon with clinical BKD. Samples collected from farm populations of rainbow trout, salmon and brown trout Salmo trutta, exposed to Rs but without clinical BKD, were negative in the ELISA, although Rs bacteria or soluble antigens were detected at the same sampling. The antibody ELISA method cannot be recommended for general fish health monitoring purposes, but may be a valuable tool for monitoring the disease progression during controlled expel-iments.

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Detection ofRenibacterium salmoninarumInfection in Asymptomatic Atlantic Salmon

Cited by 25 publications

References 11 publications

Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta

Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta

Bench‐top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture

Detection of humoral antibodies to Renibacterium salmoninarum in rainbow trout Oncorhynchus mykiss and Atlantic salmon Salmo salar challenged by immersion and in naturally infected populations

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