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Evaluation of a Commercial Real-Time PCR Kit for the Detection of Mycobacterium avium subspecies paratuberculosis in Milk

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Abstract

There are several commercial test kits for Mycobacterium avium subspecies paratuberculosis (MAP) detection, each with different advantages, disadvantages, and applications. In the present study, a real-time PCR kit targeting the unique transposon sequence ISMAP02 was evaluated. The analytical sensitivity was determined using the type strain ATCC 19698, and the specificity was validated by testing fifteen MAP isolates, thirteen non-MAP Mycobacterium isolates, and eight non-Mycobacterium isolates. Six spiking experiments were performed using raw milk and reconstituted infant milk artificially contaminated with dilutions containing 100–105 MAP cells mL−1. Sensitivity and specificity were at 100 %. The detection probabilities in raw milk and reconstituted infant milk for the samples (containing 1.4 × 101 and 1.7 × 101 MAP cell 50 mL−1) were 16.6 and 91.6 %, respectively. Thus, the tested kit yielded satisfying results to detect MAP in milk.

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Acknowledgments

This work was financially supported by the German Federal Ministry of Food and Agriculture (BMEL) through the Federal Office for Agriculture and Food (BLE), Grant Number 2812HS020. This work was a part of a doctoral programme supported by the Public Authority of Agriculture Affairs and Fish Resources (PAAF), Kuwait.

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Authors and Affiliations

  1. Public Authority of Agriculture Affairs & Fish Resources (PAAF), P.O. Box 21422, 13075, Safat, Kuwait

    Ahmad Alajmi

  2. Institute for Food Quality and Food Safety, Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany

    Ahmad Alajmi, Günter Klein, Nils. Th. Grabowski, Svenja Fohler & Amir Abdulmawjood

  3. Dairy Sciences, Institute of Veterinary Food Science, Justus-Liebig University Giessen, Ludwigstrasse 21, 35390, Giessen, Germany

    Ömer Akineden

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  1. Ahmad Alajmi
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  2. Günter Klein
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  4. Svenja Fohler
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Correspondence to Amir Abdulmawjood.

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Alajmi, A., Klein, G., Grabowski, N.T. et al. Evaluation of a Commercial Real-Time PCR Kit for the Detection of Mycobacterium avium subspecies paratuberculosis in Milk. Curr Microbiol 73, 668–675 (2016). https://doi.org/10.1007/s00284-016-1109-6

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  • DOI: https://doi.org/10.1007/s00284-016-1109-6

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