Journal article
Pooled sample testing for Bonamia ostreae: A tale of two SYBR Green real-time PCR assays
Journal of Veterinary Diagnostic Investigation, Vol.29(5), pp.752-756
2017
Abstract
Pooled testing of samples is a common laboratory practice to increase efficiency and reduce expenses. We investigated the efficacy of 2 published SYBR Green real-time PCR assays when used to detect the haplosporidian parasite Bonamia ostreae in pooled samples of infected oyster tissue. Each PCR targets a different gene within the B. ostreae genome: the actin 1 gene or the 18S rRNA gene. Tissue homogenates (150 mg) of the New Zealand flat oyster Ostrea chilensis were spiked with ~1.5 × 103 purified B. ostreae cells to create experimental pools of 3, 5, and 10. Ten positive replicates of each pool size were assayed twice with each PCR and at 2 different amounts of DNA template. The PCR targeting the actin 1 gene was unable to reproducibly detect B. ostreae in any pool size. Conversely, the 18S rRNA gene PCR could reproducibly detect B. ostreae in pools of up to 5. Using a general linear model, there was a significant difference in the number of pools that correctly detected B. ostreae between each PCR (p < 0.01) and each pool size (p < 0.01). It is likely that the single copy actin 1 gene is more likely to be diluted and not detected by pooling than the multi-copy 18S rRNA gene. Our study highlights that validation data are necessary for pooled sample testing because detection efficacy may not be comparable to individual sample testing.
Details
- Title
- Pooled sample testing for Bonamia ostreae: A tale of two SYBR Green real-time PCR assays
- Authors/Creators
- H.S. Lane (Author/Creator) - University of OtagoJ.B. Jones (Author/Creator) - University of OtagoW.L. McDonald (Author/Creator) - University of Otago
- Publication Details
- Journal of Veterinary Diagnostic Investigation, Vol.29(5), pp.752-756
- Publisher
- American Association of Veterinary Laboratory Diagnosticians
- Identifiers
- 991005540171807891
- Copyright
- © 2018 by SAGE Publications
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
UN Sustainable Development Goals (SDGs)
This output has contributed to the advancement of the following goals:
Source: InCites
Metrics
16 Record Views
InCites Highlights
These are selected metrics from InCites Benchmarking & Analytics tool, related to this output
- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.2 Marine Biology
- 3.2.1002 Bivalve Ecology
- Web Of Science research areas
- Veterinary Sciences
- ESI research areas
- Plant & Animal Science