Detection of Mycobacterium avium subsp. paratuberculosis in faeces using different procedures of pre-treatment for real-time PCR in comparison to culture

Sting, Reinhard; Hrubenja, M.; Mandl, Josef; Seemann, G.; Salditt, A.; Waibel, Sarah

doi:10.1016/j.tvjl.2013.08.033

Cited by 19 publications

(14 citation statements)

References 22 publications

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“…Detection limit of PCR in MAP cases is 10 4 organisms/gm, it is also a limitation of its use. Addition of pretreatment of fecal sample using silica membrane minicolumns and magnetic particles can enhance the detection rate of MAP by PCR (Sting et al, 2014).…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Epidemiology and Management Strategies of Johne's disease in Endemic Situations

Munir

Hassan

Muhammad³

et al. 2014

Res. j. vet. pract.

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Mycobacterium paratuberculosis the subspecies of M. avium, effects wide range of animals including domestic cattle, sheep, goats, buffaloes, camelids and wild ruminants resulting in progressive and chronic enteritis known as Johne's disease (paratuberculosis). Clinically sick animals show emaciation, diarrhea and eventually death but the risk is that mostly they don't show clinical sign still can shed bacteria in feces and milk. Organism spread in the animal body through blood and lymph nodes to multiple internal organs. It is economically very important disease in livestock because effected livestock is recommended to be culled due to high treatment costs. Etiology, host range, immunology, epidemiology, stages/ forms, clinical signs, diagnostic tools and treatment have been discussed with special reference to endemic situations. Strategies to control this disease include improved management practices, testing and culling and vaccination. Modifications in management practices is not an easy job and so is the case with testing and culling; vaccine on the other hand is the simple practice but it is not usually practiced by farmers because lack of knowledge/awareness in herdsmen and availability of vaccine.All copyrights reserved to Nexus® academic publishers

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Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Epidemiology and Management Strategies of Johne's disease in Endemic Situations

Munir

Hassan

Muhammad³

et al. 2014

Res. j. vet. pract.

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“…Thus, real-time quantitative PCR methods based on the detection of insertion sequence targets IS900 and ISMav2 were developed for rapid and sensitive detection of MAP (Ravva and Stanker, 2005; Sting et al, 2014). PCR methods can be coupled with magnetic-bead separations for cleanup and concentration of MAP DNA from complex fecal samples (Leite et al, 2013; Sting et al, 2014). PCR methods can rapidly detect the presence of MAP, but they cannot aid in typing the strains necessary for source tracking the pathogen.…”

Section: Introductionmentioning

confidence: 99%

Characterization and Differentiation of Mycobacterium avium subsp. paratuberculosis from Other Mycobacteria Using Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Ravva

Harden

Sarreal

2017

Front. Cell. Infect. Microbiol.

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Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease in cattle, is responsible for significant economic losses to the US dairy industry. The pathogen has also been associated with chronic human diseases like Crohn's disease, type 1 diabetes and multiple sclerosis. Determining causation requires rapid characterization and source tracking the pathogen. Here, we used matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to characterize and differentiate strains of MAP from 14 other species of Mycobacterium from bovine, human, and environmental sources. Lysates from cells disrupted by bead beating in TFA-acetonitrile solution were analyzed by MALDI-TOF. MAP strains were differentiated by mass spectral profiles that are distinct from each other and from other Mycobacterium species. Cluster analysis of spectral profiles indicates two distinct clusters, one dominated by the members of avium complex and a second group dominated by members of fortuitum and parafortuitum complexes. We believe that MALDI-TOF methods can be used to differentiate and source-track MAP strains.

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“…Cohen’s κ values point to a moderate accordance. qPCR B encompasses a DNA extraction protocol that includes a microfiltration step in combination with silica membrane-based mini-columns which has been shown to be one of the most sensitive protocols [ 22 ].…”

Section: Discussionmentioning

confidence: 99%

“…However, similar extraction efficiency can be achieved with optimized silica membrane-based as well as magnetic bead-based procedures [ 21 ]. The detection rate can be improved by including a microfiltration step in the sample pre-treatment [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of different diagnostic methods for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples

Hahn¹,

Failing

Eisenberg

et al. 2017

BMC Vet Res

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BackgroundEnvironmental sampling based on boot swabs and/or liquid manure samples is an upcoming strategy for the identification of paratuberculosis (paraTB) positive herds, but only limited data are available regarding the diagnostic performance of molecular detection methods (qPCR) versus faecal culture (FC) for this purpose. In the present study, the test characteristics of two different qPCR protocols (A and B) and a standardized FC protocol, for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples were evaluated.ResultsIn 19 paraTB unsuspicious and 58 paraTB positive herds boot swabs and liquid manure were sampled simultaneously and analyzed in three different diagnostic laboratories. Using boot swabs and liquid manure, a substantial to excellent accordance was found between both qPCRs, for boot swabs also with culture, while for liquid manure the detection rate of culture was decreased after prolonged storage at −20 °C. The quantitative results of both qPCR methods correlated well for the same sample and also for boot swabs and liquid manure from the same herd. When cut-off threshold cycle (CT-)-values were applied as recommended by the manufacturers, herd level specificity (Sp) of qPCR B was below 100% for boot swabs and for both qPCRs for liquid manure. A decreased herd level sensitivity was encountered after adjustment of Sp to 100% and re-calculation of the cut-off CT-values.ConclusionsqPCR is equally suitable as bacterial culture for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples. Both matrices represent easily accessible composite environmental samples which can be tested with reliable results. The data encourage qPCR testing of composite environmental samples for paraTB herd diagnosis.

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Detection of Mycobacterium avium subsp. paratuberculosis in faeces using different procedures of pre-treatment for real-time PCR in comparison to culture

Cited by 19 publications

References 22 publications

Epidemiology and Management Strategies of Johne's disease in Endemic Situations

Epidemiology and Management Strategies of Johne's disease in Endemic Situations

Characterization and Differentiation of Mycobacterium avium subsp. paratuberculosis from Other Mycobacteria Using Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Evaluation of different diagnostic methods for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples

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