Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis

Garry O. Kelley, Francisco J. Zagmutt-Vergara, Christian M. Leutenegger, Karin A. Myklebust, Mark A. Adkison, Terry S. McDowell, Gary D. Marty, Alex L. Kahler, Arla L. Bush, Ian Gardner, Ronald Hedrick

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Diagnostic methods were used to identify and quantify Myxobolus cerebralis, a myxozoan parasite of salmonid fish. In this study, 7-week-old, pathogen-free rainbow trout (Oncorhynchus mykiss) were experimentally infected with M. cerebralis and at 7 months postinfection were evaluated with 5 diagnostic assays: 1) pepsin-trypsin digest (PTD) to detect and enumerate spores found in cranial cartilage, 2) 2 different histopathology grading scales that provide a numerical score for severity of microscopic lesions in the head, 3) a conventional single-round polymerase chain reaction (PCR), 4) a nested PCR assay, and 5) a newly developed quantitative real-time TaqMan PCR. There were no significant differences (P > 0.05) among the 5 diagnostic assays in distinguishing between experimentally infected and uninfected control fish. The 2 histopathology grading scales were highly correlated (P < 0.001) for assessment of microscopic lesion severity. Quantification of parasite levels in cranial tissues using PTD and real-time TaqMan PCR was significantly correlated r = 0.540 (P < 0.001). Lastly, 104 copies of the 18S rDNA gene are present in the M. cerebralis genome, a feature that makes this gene an excellent target for PCR-based diagnostic assays. Also, 2 copies of the insulin growth factor-I gene are found in the rainbow trout genome, whose detection can serve both as an internal quality control for amplifiable DNA and as a basis to quantify pathogen genome equivalents present in quantitative PCR assays.

Original languageEnglish (US)
Pages (from-to)202-211
Number of pages10
JournalJournal of Veterinary Diagnostic Investigation
Volume16
Issue number3
DOIs
StatePublished - Jan 1 2004

Fingerprint

Myxobolus
Myxobolus cerebralis
diagnostic techniques
Oncorhynchus mykiss
Polymerase Chain Reaction
Pepsin A
assays
Genome
quantitative polymerase chain reaction
Trypsin
polymerase chain reaction
Real-Time Polymerase Chain Reaction
pepsin
Fishes
Parasites
lesions (animal)
histopathology
trypsin
genome
Genes

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Kelley, G. O., Zagmutt-Vergara, F. J., Leutenegger, C. M., Myklebust, K. A., Adkison, M. A., McDowell, T. S., ... Hedrick, R. (2004). Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis. Journal of Veterinary Diagnostic Investigation, 16(3), 202-211. https://doi.org/10.1177/104063870401600305

Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis. / Kelley, Garry O.; Zagmutt-Vergara, Francisco J.; Leutenegger, Christian M.; Myklebust, Karin A.; Adkison, Mark A.; McDowell, Terry S.; Marty, Gary D.; Kahler, Alex L.; Bush, Arla L.; Gardner, Ian; Hedrick, Ronald.

In: Journal of Veterinary Diagnostic Investigation, Vol. 16, No. 3, 01.01.2004, p. 202-211.

Research output: Contribution to journalArticle

Kelley, GO, Zagmutt-Vergara, FJ, Leutenegger, CM, Myklebust, KA, Adkison, MA, McDowell, TS, Marty, GD, Kahler, AL, Bush, AL, Gardner, I & Hedrick, R 2004, 'Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis', Journal of Veterinary Diagnostic Investigation, vol. 16, no. 3, pp. 202-211. https://doi.org/10.1177/104063870401600305
Kelley, Garry O. ; Zagmutt-Vergara, Francisco J. ; Leutenegger, Christian M. ; Myklebust, Karin A. ; Adkison, Mark A. ; McDowell, Terry S. ; Marty, Gary D. ; Kahler, Alex L. ; Bush, Arla L. ; Gardner, Ian ; Hedrick, Ronald. / Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis. In: Journal of Veterinary Diagnostic Investigation. 2004 ; Vol. 16, No. 3. pp. 202-211.
@article{3baea5cbda3849c29889b8a77b7ce587,
title = "Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis",
abstract = "Diagnostic methods were used to identify and quantify Myxobolus cerebralis, a myxozoan parasite of salmonid fish. In this study, 7-week-old, pathogen-free rainbow trout (Oncorhynchus mykiss) were experimentally infected with M. cerebralis and at 7 months postinfection were evaluated with 5 diagnostic assays: 1) pepsin-trypsin digest (PTD) to detect and enumerate spores found in cranial cartilage, 2) 2 different histopathology grading scales that provide a numerical score for severity of microscopic lesions in the head, 3) a conventional single-round polymerase chain reaction (PCR), 4) a nested PCR assay, and 5) a newly developed quantitative real-time TaqMan PCR. There were no significant differences (P > 0.05) among the 5 diagnostic assays in distinguishing between experimentally infected and uninfected control fish. The 2 histopathology grading scales were highly correlated (P < 0.001) for assessment of microscopic lesion severity. Quantification of parasite levels in cranial tissues using PTD and real-time TaqMan PCR was significantly correlated r = 0.540 (P < 0.001). Lastly, 104 copies of the 18S rDNA gene are present in the M. cerebralis genome, a feature that makes this gene an excellent target for PCR-based diagnostic assays. Also, 2 copies of the insulin growth factor-I gene are found in the rainbow trout genome, whose detection can serve both as an internal quality control for amplifiable DNA and as a basis to quantify pathogen genome equivalents present in quantitative PCR assays.",
author = "Kelley, {Garry O.} and Zagmutt-Vergara, {Francisco J.} and Leutenegger, {Christian M.} and Myklebust, {Karin A.} and Adkison, {Mark A.} and McDowell, {Terry S.} and Marty, {Gary D.} and Kahler, {Alex L.} and Bush, {Arla L.} and Ian Gardner and Ronald Hedrick",
year = "2004",
month = "1",
day = "1",
doi = "10.1177/104063870401600305",
language = "English (US)",
volume = "16",
pages = "202--211",
journal = "Journal of Veterinary Diagnostic Investigation",
issn = "1040-6387",
publisher = "American Association of Veterinary Laboratory Diagnosticians",
number = "3",

}

TY - JOUR

T1 - Evaluation of five diagnostic methods for the detection and quantification of Myxobolus cerebralis

AU - Kelley, Garry O.

AU - Zagmutt-Vergara, Francisco J.

AU - Leutenegger, Christian M.

AU - Myklebust, Karin A.

AU - Adkison, Mark A.

AU - McDowell, Terry S.

AU - Marty, Gary D.

AU - Kahler, Alex L.

AU - Bush, Arla L.

AU - Gardner, Ian

AU - Hedrick, Ronald

PY - 2004/1/1

Y1 - 2004/1/1

N2 - Diagnostic methods were used to identify and quantify Myxobolus cerebralis, a myxozoan parasite of salmonid fish. In this study, 7-week-old, pathogen-free rainbow trout (Oncorhynchus mykiss) were experimentally infected with M. cerebralis and at 7 months postinfection were evaluated with 5 diagnostic assays: 1) pepsin-trypsin digest (PTD) to detect and enumerate spores found in cranial cartilage, 2) 2 different histopathology grading scales that provide a numerical score for severity of microscopic lesions in the head, 3) a conventional single-round polymerase chain reaction (PCR), 4) a nested PCR assay, and 5) a newly developed quantitative real-time TaqMan PCR. There were no significant differences (P > 0.05) among the 5 diagnostic assays in distinguishing between experimentally infected and uninfected control fish. The 2 histopathology grading scales were highly correlated (P < 0.001) for assessment of microscopic lesion severity. Quantification of parasite levels in cranial tissues using PTD and real-time TaqMan PCR was significantly correlated r = 0.540 (P < 0.001). Lastly, 104 copies of the 18S rDNA gene are present in the M. cerebralis genome, a feature that makes this gene an excellent target for PCR-based diagnostic assays. Also, 2 copies of the insulin growth factor-I gene are found in the rainbow trout genome, whose detection can serve both as an internal quality control for amplifiable DNA and as a basis to quantify pathogen genome equivalents present in quantitative PCR assays.

AB - Diagnostic methods were used to identify and quantify Myxobolus cerebralis, a myxozoan parasite of salmonid fish. In this study, 7-week-old, pathogen-free rainbow trout (Oncorhynchus mykiss) were experimentally infected with M. cerebralis and at 7 months postinfection were evaluated with 5 diagnostic assays: 1) pepsin-trypsin digest (PTD) to detect and enumerate spores found in cranial cartilage, 2) 2 different histopathology grading scales that provide a numerical score for severity of microscopic lesions in the head, 3) a conventional single-round polymerase chain reaction (PCR), 4) a nested PCR assay, and 5) a newly developed quantitative real-time TaqMan PCR. There were no significant differences (P > 0.05) among the 5 diagnostic assays in distinguishing between experimentally infected and uninfected control fish. The 2 histopathology grading scales were highly correlated (P < 0.001) for assessment of microscopic lesion severity. Quantification of parasite levels in cranial tissues using PTD and real-time TaqMan PCR was significantly correlated r = 0.540 (P < 0.001). Lastly, 104 copies of the 18S rDNA gene are present in the M. cerebralis genome, a feature that makes this gene an excellent target for PCR-based diagnostic assays. Also, 2 copies of the insulin growth factor-I gene are found in the rainbow trout genome, whose detection can serve both as an internal quality control for amplifiable DNA and as a basis to quantify pathogen genome equivalents present in quantitative PCR assays.

UR - http://www.scopus.com/inward/record.url?scp=2442621210&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442621210&partnerID=8YFLogxK

U2 - 10.1177/104063870401600305

DO - 10.1177/104063870401600305

M3 - Article

C2 - 15152834

AN - SCOPUS:2442621210

VL - 16

SP - 202

EP - 211

JO - Journal of Veterinary Diagnostic Investigation

JF - Journal of Veterinary Diagnostic Investigation

SN - 1040-6387

IS - 3

ER -