TitleDevelopment of Quantitative Real-Time PCR Assays for Postmortem Detection of spp. Common in Zebrafish () Research Colonies.
Publication TypeJournal Article
Year of Publication2017
AuthorsMeritet, DM, Mulrooney, DM, Kent, ML, Löhr, CV
JournalJ Am Assoc Lab Anim Sci
Volume56
Issue2
Pagination131-141
Date Published2017 Mar 01
ISSN1559-6109
KeywordsAnimals, Fish Diseases, Laboratory Animal Science, Mycobacterium, Mycobacterium Infections, Real-Time Polymerase Chain Reaction, Zebrafish
Abstract

Mycobacterium spp. infections are common in zebrafish kept in research facilities. These comorbidities can substantially modulate the responses of these fish to external and internal stimuli. Therefore, diagnostic tests to detect Mycobacterium spp. infections in zebrafish colonies prove essential. Here, we outline the development of quantitative simplex real-time PCR assays to detect the 3 Mycobacterium species most commonly identified in laboratory zebrafish. The assays targeted the heatshock protein 65 gene of M. marinum, M. chelonae, and M. haemophilum. The assays are both highly specific and sensitive for fresh-frozen samples and highly specific and moderately sensitive for formalin-fixed paraffin-embedded (FFPE) samples. Two sampling techniques for FFPE samples of sagittally sectioned zebrafish were evaluated. Both paraffin cores targeting granulomas containing bacteria and scrolls from the entire fish yielded DNA of equivalent quantity and purity. The diagnostic sensitivity of cores was superior to that of scrolls for M. chelonae and M. haemophilum but not M. marinum. The assays are cost-effective and ideally suited to diagnosing common Mycobacterium spp. infections in zebrafish.

Alternate JournalJ. Am. Assoc. Lab. Anim. Sci.
PubMed ID28315641
PubMed Central IDPMC5361037
Grant ListP30 ES000210 / ES / NIEHS NIH HHS / United States
P40 OD011021 / OD / NIH HHS / United States
R24 OD010998 / OD / NIH HHS / United States
T32 RR023917 / RR / NCRR NIH HHS / United States