Validation of IS900- qPCR assay to assess the presence of Mycobacterium avium subs. paratuberculosis in faecal samples according to the OIE procedure

Russo, Simone; Giorgio, Galletti; Leo, Simone; Arrigoni, N.; Garbarino, C.; Ricchi, M.

doi:10.1016/j.prevetmed.2022.105732

Cited by 8 publications

(7 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using a Bayesian latent class model, median diagnostic specificity of an IS900 PCR assay to detect MAP DNA in bovine fecal samples has been estimated to be ~96% for prevalence scenarios of 20-70%. 21 The high specificity of ISMap02 that we observed might be explained by the specificity of the target gene. In a previous study, PCR methods using ISMap02 did not amplify DNA of any of the genetically close mycobacteria.…”

Section: Discussionmentioning

confidence: 82%

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis

Arango-Sabogal,

Labrecque,

Fairbrother

et al. 2023

J VET Diagn Invest

View full text Add to dashboard Cite

Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of paratuberculosis, a chronic, contagious, and incurable enteric disease of ruminants. An in-house IS 900 PCR assay validated for MAP detection in sheep has been shown to have a higher sensitivity than a commercial PCR and fecal culture. We have now compared the performance of this in-house IS 900 PCR assay with a commercial ISMap 02 PCR assay for the detection of MAP DNA in bovine dairy farm environmental samples. We purposefully selected 30 culture-positive, 62 culture-negative, and 62 non-interpretable environmental samples. We applied the IS 900 PCR assay directly to the frozen inoculum of these samples. Inocula were incubated in an automated system, and growth was confirmed by an acid-fast bacilli stain and the IS 900 PCR assay. Among culture-positive samples before incubation, the IS 900 PCR assay yielded significantly more positive results than the ISMap 02 PCR assay; however, among culture-negative samples, the IS 900 PCR assay yielded positive results both before and after incubation. The ISMap 02 PCR assay did not flag positively among the culture-negative samples either before or after incubation. The IS 900 PCR assay is a sensitive method that can be used to detect MAP DNA in environmental samples before incubation. The ISMap 02 PCR assay is a specific method used to detect MAP DNA in environmental samples both before and after incubation.

show abstract

Section: Discussionmentioning

confidence: 82%

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis

Arango-Sabogal,

Labrecque,

Fairbrother

et al. 2023

J VET Diagn Invest

View full text Add to dashboard Cite

show abstract

“…Additionally, tests were performed at the discretion of the official veterinarian based on farm health status. The procedure consisted of probe‐based qPCR targeting the IS900 sequence used for the detection of MAP DNA, as previously described 14,15 …”

Section: Methodsmentioning

confidence: 99%

“…The procedure consisted of probe-based qPCR targeting the IS900 sequence used for the detection of MAP DNA, as previously described. 14,15…”

Section: Indirect and Direct Diagnosismentioning

confidence: 99%

Long‐term outcomes of the Italian Mycobacterium avium subspecies paratuberculosis control programme for dairy cattle

Musolino,

Rampacci,

Tolasi

et al. 2024

Veterinary Record

View full text Add to dashboard Cite

BackgroundThe considerable epidemiological and economic implications of paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), have placed importance on control efforts aimed at preventing MAP transmission. In this context, Italy issued national guidelines for the control and status certification of MAP in dairy cattle in 2013.MethodsWe assessed the long‐term outcomes of the Italian MAP control programme for 14 dairy farms located in northern Italy by retrospectively reviewing the results of yearly serological tests, presence of clinical cases, MAP faecal shedding in serologically positive animals, farm management and health ranking as indicators of herd health between 2014 and 2021.ResultsA significantly higher number of serologically positive animals were observed between 2014 and 2016 than between 2017 and 2021, as well as an improving trend in the paratuberculosis health ranking for nine of the 14 farms. No clinical cases were reported. MAP shedding was detected in 9.4% of serologically positive animals. Discarding colostrum and prioritised culling of seropositive animals assisted by adoption of standardised serological testing were presumed to have a key role in MAP control, despite the reluctance of some farmers to address hygienic issues and improve the separation of calves from adult animals.LimitationsThe small number of farms included in this study and the fact that these were not randomly selected may limit the generalisability of the findings.ConclusionsThe Italian paratuberculosis control plan has provided measures to limit the uncontrolled spread of MAP infection within and between herds by promoting animal trading between farms certified as negative or low risk.

show abstract

“…To detect MAP-positive animals from tissue and feces, DNA was extracted from 25 mg of individual pools of ileocecal valves and mesenteric lymph-nodes ( 11 ) and from 3 ± 0.5 g of feces, as reported in Russo et al ( 20 ). Quantitative PCR was performed on eluted DNA targeting IS900 sequence ( 20 ). Results were expressed by cycle of quantification (Cq) values, considering Cq values of two replicates of the same sample.…”

Section: Methodsmentioning

confidence: 99%

Active surveillance of paratuberculosis in Alpine-dwelling red deer (Cervus elaphus)

Filippi,

Garbarino,

Nava

et al. 2024

Front. Vet. Sci.

Self Cite

View full text Add to dashboard Cite

Paratuberculosis (Johne’s disease) is a globally widespread infectious disease affecting domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). The bacterium is excreted in the feces and is characterized by high environmental resistance. The new Animal Health Law (Regulation EU 2016/429) on transmissible animal diseases, recently in force throughout the European Union, includes paratuberculosis within the diseases requiring surveillance in the EU, listing some domestic and wild Bovidae, Cervidae, and Camelidae as potential reservoirs. Taking advantage of a culling activity conducted in the Stelvio National Park (Italy), this study investigated MAP infection status of red deer (Cervus elaphus) between 2018 and 2022, and evaluated the probability of being MAP-positive with respect to individual and sampling-level variables. A total of 390 subjects were examined macroscopically and tested for MAP, using different diagnostic tools: IS900 qPCR, culture, histopathology, and serology. Twenty-three of them were found positive for MAP by at least one test, with an overall prevalence of 5.9% (95% CI 4.0–8.7), that, respectively, ranged from 12.4% in the first culling season to 2.0 and 2.1% in the 2019–2020 and 2021–2022 culling seasons. Quantitative PCR assay on ileocecal valve and mesenteric lymph nodes detected the highest number of MAP positive animals. The results of the study showed the increased probability of being MAP-positive with increasing age and that red deer with lower body mass values were more likely to be infected with MAP. Overall, the absence of signs of clinical paratuberculosis and gross lesions together with the low level of shedding witness early phases of the disease among the positive red deer and support an improvement of the paratuberculosis status of this population, as shown by the decreased prevalence of the disease over the years.

show abstract

Validation of IS900- qPCR assay to assess the presence of Mycobacterium avium subs. paratuberculosis in faecal samples according to the OIE procedure

Cited by 8 publications

References 37 publications

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis

Long‐term outcomes of the Italian Mycobacterium avium subspecies paratuberculosis control programme for dairy cattle

Active surveillance of paratuberculosis in Alpine-dwelling red deer (Cervus elaphus)

Contact Info

Product

Resources

About