PCR and probe-PCR assays to monitor broodstock Atlantic salmon (Salmo salar L.) ovarian fluid and kidney tissue for presence of DNA of the fish pathogen Renibacterium salmoninarum
A simple, rapid PCR assay for the identification of Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.) tissues detected DNA extracted from between 4 and 40 bacterial cells. PCR was at least as sensitive as culture when it was used to identify subclinically infected fish experimentally ch...
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| Published in: | Journal of clinical microbiology Vol. 35; no. 6; p. 1322 |
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| Main Authors: | , , , |
| Format: | Journal Article |
| Language: | English |
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United States
01.06.1997
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| ISSN: | 0095-1137 |
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| Abstract | A simple, rapid PCR assay for the identification of Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.) tissues detected DNA extracted from between 4 and 40 bacterial cells. PCR was at least as sensitive as culture when it was used to identify subclinically infected fish experimentally challenged with R. salmoninarum. However, PCR identified much higher numbers of kidney tissue and ovarian fluid samples from commercially reared broodstock fish to be positive for R. salmoninarum than did culture. This difference may be due to the antibiotic chemotherapy of broodstock fish used by the industry in 1994 to control the vertical transmission of R. salmoninarum. A much closer relationship between PCR and culture results was observed for ovarian fluid samples collected from broodstock fish in 1993. Also, PCR scored a much higher percentage of kidney tissue samples than ovarian fluid samples from 1994 broodstock fish positive for R. salmoninarum, which may reflect the uneven distribution of the pathogen in different fish tissues. Inclusion of a nested probe to identify the PCR-positive 1994 ovarian fluid samples increased the sensitivity of detection to between one and four cells and the number of samples that scored positive by almost threefold. These data indicate that many infected ovarian fluid samples contained very low numbers of R. salmoninarum cells and, because almost all these samples were culture negative, that PCR may have detected dead or otherwise unculturable bacterial cells. |
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| AbstractList | A simple, rapid PCR assay for the identification of Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.) tissues detected DNA extracted from between 4 and 40 bacterial cells. PCR was at least as sensitive as culture when it was used to identify subclinically infected fish experimentally challenged with R. salmoninarum. However, PCR identified much higher numbers of kidney tissue and ovarian fluid samples from commercially reared broodstock fish to be positive for R. salmoninarum than did culture. This difference may be due to the antibiotic chemotherapy of broodstock fish used by the industry in 1994 to control the vertical transmission of R. salmoninarum. A much closer relationship between PCR and culture results was observed for ovarian fluid samples collected from broodstock fish in 1993. Also, PCR scored a much higher percentage of kidney tissue samples than ovarian fluid samples from 1994 broodstock fish positive for R. salmoninarum, which may reflect the uneven distribution of the pathogen in different fish tissues. Inclusion of a nested probe to identify the PCR-positive 1994 ovarian fluid samples increased the sensitivity of detection to between one and four cells and the number of samples that scored positive by almost threefold. These data indicate that many infected ovarian fluid samples contained very low numbers of R. salmoninarum cells and, because almost all these samples were culture negative, that PCR may have detected dead or otherwise unculturable bacterial cells.A simple, rapid PCR assay for the identification of Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.) tissues detected DNA extracted from between 4 and 40 bacterial cells. PCR was at least as sensitive as culture when it was used to identify subclinically infected fish experimentally challenged with R. salmoninarum. However, PCR identified much higher numbers of kidney tissue and ovarian fluid samples from commercially reared broodstock fish to be positive for R. salmoninarum than did culture. This difference may be due to the antibiotic chemotherapy of broodstock fish used by the industry in 1994 to control the vertical transmission of R. salmoninarum. A much closer relationship between PCR and culture results was observed for ovarian fluid samples collected from broodstock fish in 1993. Also, PCR scored a much higher percentage of kidney tissue samples than ovarian fluid samples from 1994 broodstock fish positive for R. salmoninarum, which may reflect the uneven distribution of the pathogen in different fish tissues. Inclusion of a nested probe to identify the PCR-positive 1994 ovarian fluid samples increased the sensitivity of detection to between one and four cells and the number of samples that scored positive by almost threefold. These data indicate that many infected ovarian fluid samples contained very low numbers of R. salmoninarum cells and, because almost all these samples were culture negative, that PCR may have detected dead or otherwise unculturable bacterial cells. A simple, rapid PCR assay for the identification of Renibacterium salmoninarum in Atlantic salmon (Salmo salar L.) tissues detected DNA extracted from between 4 and 40 bacterial cells. PCR was at least as sensitive as culture when it was used to identify subclinically infected fish experimentally challenged with R. salmoninarum. However, PCR identified much higher numbers of kidney tissue and ovarian fluid samples from commercially reared broodstock fish to be positive for R. salmoninarum than did culture. This difference may be due to the antibiotic chemotherapy of broodstock fish used by the industry in 1994 to control the vertical transmission of R. salmoninarum. A much closer relationship between PCR and culture results was observed for ovarian fluid samples collected from broodstock fish in 1993. Also, PCR scored a much higher percentage of kidney tissue samples than ovarian fluid samples from 1994 broodstock fish positive for R. salmoninarum, which may reflect the uneven distribution of the pathogen in different fish tissues. Inclusion of a nested probe to identify the PCR-positive 1994 ovarian fluid samples increased the sensitivity of detection to between one and four cells and the number of samples that scored positive by almost threefold. These data indicate that many infected ovarian fluid samples contained very low numbers of R. salmoninarum cells and, because almost all these samples were culture negative, that PCR may have detected dead or otherwise unculturable bacterial cells. |
| Author | Lynch, W H Lovely, J E Miriam, A Griffiths, S G |
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| References | 3524440 - Antimicrob Agents Chemother. 1986 Jun;29(6):992-6 1383085 - FEMS Microbiol Lett. 1992 Sep 15;75(2-3):259-65 2686828 - Can J Vet Res. 1989 Oct;53(4):385-9 8138127 - FEMS Microbiol Lett. 1994 Jan 15;115(2-3):131-6 7592363 - J Bacteriol. 1995 Nov;177(21):6027-32 7529017 - Appl Environ Microbiol. 1994 Dec;60(12):4580-3 8250575 - Appl Environ Microbiol. 1993 Oct;59(10):3513-5 3312987 - Microbiol Rev. 1987 Sep;51(3):365-79 8179886 - Biotechniques. 1994 Feb;16(2):242-4, 246 |
| References_xml | – reference: 2686828 - Can J Vet Res. 1989 Oct;53(4):385-9 – reference: 7529017 - Appl Environ Microbiol. 1994 Dec;60(12):4580-3 – reference: 8138127 - FEMS Microbiol Lett. 1994 Jan 15;115(2-3):131-6 – reference: 1383085 - FEMS Microbiol Lett. 1992 Sep 15;75(2-3):259-65 – reference: 3312987 - Microbiol Rev. 1987 Sep;51(3):365-79 – reference: 3524440 - Antimicrob Agents Chemother. 1986 Jun;29(6):992-6 – reference: 8250575 - Appl Environ Microbiol. 1993 Oct;59(10):3513-5 – reference: 7592363 - J Bacteriol. 1995 Nov;177(21):6027-32 – reference: 8179886 - Biotechniques. 1994 Feb;16(2):242-4, 246 |
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| SubjectTerms | Animals Bacteriological Techniques DNA, Bacterial - analysis Female Fish Diseases - diagnosis Fish Diseases - microbiology Gram-Positive Asporogenous Rods - genetics Gram-Positive Asporogenous Rods - isolation & purification Gram-Positive Bacterial Infections - diagnosis Gram-Positive Bacterial Infections - microbiology Gram-Positive Bacterial Infections - veterinary Kidney - microbiology Kidney Diseases - diagnosis Kidney Diseases - microbiology Kidney Diseases - veterinary Ovary - microbiology Polymerase Chain Reaction - methods Salmon - microbiology Sensitivity and Specificity |
| Title | PCR and probe-PCR assays to monitor broodstock Atlantic salmon (Salmo salar L.) ovarian fluid and kidney tissue for presence of DNA of the fish pathogen Renibacterium salmoninarum |
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