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TaqMan real-time polymerase chain reaction for detection of Ophidiomyces ophiodiicola, the fungus associated with snake fungal disease.
BMC Vet Res. 2015 Apr 15; 11:95.BV

Abstract

BACKGROUND

Fungal skin infections associated with Ophidiomyces ophiodiicola, a member of the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) complex, have been linked to an increasing number of cases of snake fungal disease (SFD) in captive snakes around the world and in wild snake populations in eastern North America. The emergence of SFD in both captive and wild situations has led to an increased need for tools to better diagnose and study the disease.

RESULTS

We developed two TaqMan real-time polymerase chain reaction (PCR) assays to rapidly detect O. ophiodiicola in clinical samples. One assay targets the internal transcribed spacer region (ITS) of the fungal genome while the other targets the more variable intergenic spacer region (IGS). The PCR assays were qualified using skin samples collected from 50 snakes for which O. ophiodiicola had been previously detected by culture, 20 snakes with gross skin lesions suggestive of SFD but which were culture-negative for O. ophiodiicola, and 16 snakes with no clinical signs of infection. Both assays performed equivalently and proved to be more sensitive than traditional culture methods, detecting O. ophiodiicola in 98% of the culture-positive samples and in 40% of the culture-negative snakes that had clinical signs of SFD. In addition, the assays did not cross-react with a panel of 28 fungal species that are closely related to O. ophiodiicola or that commonly occur on the skin of snakes. The assays did, however, indicate that some asymptomatic snakes (~6%) may harbor low levels of the fungus, and that PCR should be paired with histology when a definitive diagnosis is required.

CONCLUSIONS

These assays represent the first published methods to detect O. ophiodiicola by real-time PCR. The ITS assay has great utility for assisting with SFD diagnoses whereas the IGS assay offers a valuable tool for research-based applications.

Authors+Show Affiliations

United States Geological Survey, National Wildlife Health Center, Madison, WI, USA. ebohuski@usgs.gov.Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA. jmlorch@wisc.edu.United States Geological Survey, National Wildlife Health Center, Madison, WI, USA. kgriffin@usgs.gov.United States Geological Survey, National Wildlife Health Center, Madison, WI, USA. dblehert@usgs.gov.

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

25889462

Citation

Bohuski, Elizabeth, et al. "TaqMan Real-time Polymerase Chain Reaction for Detection of Ophidiomyces Ophiodiicola, the Fungus Associated With Snake Fungal Disease." BMC Veterinary Research, vol. 11, 2015, p. 95.
Bohuski E, Lorch JM, Griffin KM, et al. TaqMan real-time polymerase chain reaction for detection of Ophidiomyces ophiodiicola, the fungus associated with snake fungal disease. BMC Vet Res. 2015;11:95.
Bohuski, E., Lorch, J. M., Griffin, K. M., & Blehert, D. S. (2015). TaqMan real-time polymerase chain reaction for detection of Ophidiomyces ophiodiicola, the fungus associated with snake fungal disease. BMC Veterinary Research, 11, 95. https://doi.org/10.1186/s12917-015-0407-8
Bohuski E, et al. TaqMan Real-time Polymerase Chain Reaction for Detection of Ophidiomyces Ophiodiicola, the Fungus Associated With Snake Fungal Disease. BMC Vet Res. 2015 Apr 15;11:95. PubMed PMID: 25889462.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - TaqMan real-time polymerase chain reaction for detection of Ophidiomyces ophiodiicola, the fungus associated with snake fungal disease. AU - Bohuski,Elizabeth, AU - Lorch,Jeffrey M, AU - Griffin,Kathryn M, AU - Blehert,David S, Y1 - 2015/04/15/ PY - 2014/09/05/received PY - 2015/04/02/accepted PY - 2015/4/19/entrez PY - 2015/4/19/pubmed PY - 2016/1/23/medline SP - 95 EP - 95 JF - BMC veterinary research JO - BMC Vet. Res. VL - 11 N2 - BACKGROUND: Fungal skin infections associated with Ophidiomyces ophiodiicola, a member of the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) complex, have been linked to an increasing number of cases of snake fungal disease (SFD) in captive snakes around the world and in wild snake populations in eastern North America. The emergence of SFD in both captive and wild situations has led to an increased need for tools to better diagnose and study the disease. RESULTS: We developed two TaqMan real-time polymerase chain reaction (PCR) assays to rapidly detect O. ophiodiicola in clinical samples. One assay targets the internal transcribed spacer region (ITS) of the fungal genome while the other targets the more variable intergenic spacer region (IGS). The PCR assays were qualified using skin samples collected from 50 snakes for which O. ophiodiicola had been previously detected by culture, 20 snakes with gross skin lesions suggestive of SFD but which were culture-negative for O. ophiodiicola, and 16 snakes with no clinical signs of infection. Both assays performed equivalently and proved to be more sensitive than traditional culture methods, detecting O. ophiodiicola in 98% of the culture-positive samples and in 40% of the culture-negative snakes that had clinical signs of SFD. In addition, the assays did not cross-react with a panel of 28 fungal species that are closely related to O. ophiodiicola or that commonly occur on the skin of snakes. The assays did, however, indicate that some asymptomatic snakes (~6%) may harbor low levels of the fungus, and that PCR should be paired with histology when a definitive diagnosis is required. CONCLUSIONS: These assays represent the first published methods to detect O. ophiodiicola by real-time PCR. The ITS assay has great utility for assisting with SFD diagnoses whereas the IGS assay offers a valuable tool for research-based applications. SN - 1746-6148 UR - https://www.unboundmedicine.com/medline/citation/25889462/TaqMan_real_time_polymerase_chain_reaction_for_detection_of_Ophidiomyces_ophiodiicola_the_fungus_associated_with_snake_fungal_disease_ L2 - https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-015-0407-8 DB - PRIME DP - Unbound Medicine ER -