FEB 04, 2015 06:00 AM PST

PCR testing in rodent health monitoring program: where are we exactly?

  • Assistant Director of the Animal Resources Department, Salk Institute for Biological Studies
      Dr. Leblanc is the Assistant Director of the Animal Resources Department at the Salk Institute for Biological studies. He obtained his Doctorate in Veterinary Medicine from the University of Montreal in 1997. After 2 years in private practice, he pursued a Masters and a Ph.D. degree in Physiology and Endocrinology. During his doctoral studies, Dr. Leblanc characterized the pharmacokinetic and peripheral metabolism of DHEA in cynolmogus monkeys and also participated in the validation of human Affymetrix genechip microarrays in non-human primates for genomic studies. While doing his doctoral studies, he developed a unique expertise in Laboratory Animal Medicine and Drug Safety assessment in both the biotechnology and academic sectors. Following his Ph.D., he did a residency in Veterinary Pathology at the University of Montreal while remaining active in Lab Animal Science. He then became Director of Regulatory Affairs and University Veterinarian at McGill University overseeing one of the largest animal care and use program in Canada. He moved on to the position of Assistant Director and Veterinary Pathologist at the Salk Institute in 2007 where he is responsible for the veterinary care, diagnostic and preventative medicine program as well as the Comparative Pathology platform. Dr. Leblanc is also a Staff Scientist in the Gene Expression Laboratory and has published on cancer research and metabolism in several high impact journals such as Cell, Cancer Cell and Nature Medicine. He is Diplomate of the ACLAM, participated on multiple assessment visits for both the Canadian Council on Animal Care (CCAC) and AAALAC International and published several articles in Lab Animal Medicine.
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    Soiled-bedding sentinels are not always efficient in detecting pathogens in rodent colonies. In this context, PCR-based testing can be more sensitive and is being advocated as adjunct to traditional health-monitoring methods. PCR testing of biological samples is commonly used to confirm positive results from sentinels or detect pathogens in quarantine. Intra-cage environmental PCR is sensitive in detecting some agents but requires large sampling sizes to be representative and can be costly and unrewarding, especially when pathogen prevalence is low. Another strategy is to sample exhaust air dust from ventilated racks where particles accumulate to detect pathogens at the rack level. This strategy has proven useful in detecting fur mites, MHV and Sendai virus, but results were less conclusive for other agents. Overall, optimal sampling strategies for environmental PCR have not been established for different microorganisms. As importantly, PCR testing may yield false-positives by amplifying nonspecific DNA or identifying genetic material in the absence of infectious particles. Our group was one of the first to raise concerns on environmental PCR following false-positive pinworm results due to non-specificity of commercial assays and amplification of Rhabditid nematodes in bedding. In this presentation, we will build on our past experience with environmental PCR in dealing with pathogen outbreaks and will describe current strategies on these new diagnostic assays. We will review the methodology used by reference laboratories to validate PCR assays and discuss the necessary controls to integrate these molecular methods efficiently and economically in rodent health monitoring programs.

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