Worsley, L and Davies, PL 
ORCID: 0000-0001-6085-9763
(2024)
Inter-laboratory ring trial to compare four quantitative polymerase chain reaction assays employed for detection of <i>Mycobacterium avium</i> subspecies <i>paratuberculosis</i>.
Microbiology spectrum, 12 (3).
e0221023-e0221023.
Abstract
Johne's disease is an infectious enteric disease caused by <i>Mycobacterium avium</i> subspecies <i>paratuberculosi</i>s (MAP) affecting ruminant species worldwide. In Project 1, an independent performance comparison ring trail was conducted between three different commercial MAP quantitative polymerase chain reaction (qPCR) assay services (B, C, and D) currently marketed in Great Britain by three separate laboratories against each other and against a fourth assay (A) not available commercially in Great Britain. A total of 205 individual ovine and bovine samples from five farms were analyzed to give 41 sets of pooled results (pool size five) from each laboratory according to their specific protocols. The numbers of positive pools for assays A-D were 18, 12, 11, and 1 (43.9%, 29.2%, 26.8%, and 2.4%), respectively. Assessment of interrater reliability produced a Fleiss' kappa coefficient of 0.15, indicating very poor overall agreement between the four laboratories. Laboratories A-D diagnosed 4, 3, 2, and 1 flocks at the farm level, respectively, as MAP positive. In Project 2, 38 pooled ovine samples from 10 flocks were analyzed to compare the performance of laboratories A and B. The numbers of positive results for laboratories A and B were 24 (63.1%) and 17 (44.7%), respectively (Cohen's kappa 0.54), indicating that laboratory A was more sensitive than B in line with results from Project 1. Variation between laboratories offering MAP qPCR assays is a significant concern, and further work is warranted to validate and standardize the performance of assays between laboratories for both ovine and bovine samples.IMPORTANCEOur study reports the findings of an inter-laboratory ring trial comparing the performance of four different quantitative polymerase chain reaction (qPCR) assay services for detecting <i>Mycobacterium avium</i> subspecies <i>paratuberculosis</i> (MAP) infection in cattle and sheep. MAP is the causative agent of Johne's disease (also known as paratuberculosis), a significant production-limiting disease in livestock populations with a worldwide distribution. The content of this paper is significant and novel as it is the first to highlight the marked variation between the diagnostic sensitivity and reproducibility of the three principal commercial laboratories offering MAP qPCR diagnostic and screening services in Great Britain. The low sensitivity and high variability between the laboratories are of great concern and relevance to veterinary practitioners and livestock producers.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Feces, Animals, Cattle, Sheep, Paratuberculosis, Cattle Diseases, Reproducibility of Results, Polymerase Chain Reaction, Mycobacterium avium subsp. paratuberculosis |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 26 Mar 2024 10:55 |
| Last Modified: | 26 Mar 2024 15:35 |
| DOI: | 10.1128/spectrum.02210-23 |
| Open Access URL: | https://doi.org/10.1128/spectrum.02210-23 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3179962 |
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