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Journal of Veterinary Diagnostic Investigation Vol. 22 Issue 1, 90-93
Copyright © 2010 by the American Association of Veterinary Laboratory Diagnosticians
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Brief Research Reports

In situ hybridization for the detection of Mycoplasma bovis in paraffin-embedded lung tissue from experimentally infected calves

Björn Jacobsen, Kathrin Hermeyer, Wolfgang Jechlinger{dagger}, Martina Zimmermann, Joachim Spergser, Renate Rosengarten and Marion Hewicker-Trautwein1

Correspondence: 1Corresponding Author: Marion Hewicker-Trautwein, Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, D-30559 Hannover, Germany, e-mail: Marion.Hewicker-Trautwein{at}tiho-hannover.de

Mycoplasma bovis DNA was detected in lung tissue of experimentally infected calves by in situ hybridization (ISH) with a nonradioactive, digoxigenin-labeled DNA probe. The 171–base pair DNA probe targeting part of the gene of the major immunodominant variable surface protein A, which is conserved among all vsp genes, was generated by polymerase chain reaction. Four calves between 57 and 63 days old were inoculated intratracheally with 30 ml of a suspension of M. bovis strain 1067 containing 7 x 104 colony forming units per milliliter. Two calves inoculated with 30 ml of sterile medium served as control animals. The calves were euthanized and then examined 21 days after inoculation. The ISH method developed in the current study was suitable for the detection of M. bovis DNA in formalin-fixed, paraffin-embedded lung tissue and may be a valuable tool for diagnostic purposes and for further investigating the pathogenesis of M. bovis infection.

Key Words: Cattle • in situ hybridization • lung • Mycoplasma bovis






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