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Tierarztl Prax Ausg G Grosstiere Nutztiere 2010; 38(06): 357-362
DOI: 10.1055/s-0038-1624004
Original Article
Schattauer GmbH

Immunohistochemistry and polymerase chain reaction for detection of Mycoplasma hyopneumoniae infection in piglets

Vergleich von Immunhistochemie und Polymerasekettenreaktion zum Nachweis von M. -hyopneumoniae-Infektionen bei jungen Ferkeln
L. Moorkamp
1   Field Station for Epidemiology, University of Veterinary Medicine Hannover, Foundation
,
E. grosse Beilage
1   Field Station for Epidemiology, University of Veterinary Medicine Hannover, Foundation
,
M. Hewicker-Trautwein
2   Institute for Pathology, University of Veterinary Medicine Hannover, Foundation
› Author Affiliations
Further Information

Publication History

Received: 25 July 2010

Accepted after revision: 30 September 2010

Publication Date:
06 January 2018 (online)

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Summary:

Objective: Few studies exist that concentrate on the detection of M. hyopneumoniae by PCR in piglets. Most of these studies do not permit the differentiation of latent infections from infections with typical pathomorphological lesions. The aim of the study was to characterize the occurrence of M. hyopneumoniae infections in piglets by combining and evaluating the results of macroscopical, histological, molecularbiological and immunohistochemical examinations. The evaluation of the suitability of the immunohistochemistry (IHC) as an alternative or as a supplement to other tests was a further aim. The study-design does not allow any conclusions regarding the prevalence of early M. hyo - pneumoniae-infections in piglets. Material and methods: 96 clinically affected piglets from eight pre-selected herds with an increased risk for M. hyopneumoniae infection were chosen for necropsy. Macroscopical and histological examinations of the lungs were performed. BALF and lung tissue were taken for the detection of M. hyopneumoniae by nested PCR (nPCR) and IHC. Results: 16 of 33 nPCR-positive piglets were positive for antigen by IHC while 63 nPCR-negative piglets were also negative for antigen by IHC (kappa index = 0.55). Of the 17 piglets with a nPCR-positive but IHC-negative test result, 12 originated from one herd leading to the assumption that not all field strains were detectable with the monoclonal antibody utilized. Fisher’s exact test showed a significant association (p < 0.0001) between detection of M. hyopneumoniae by nPCR in BALF or by IHC in lung tissue and the occurrence of typical pathological lesions. Conclusion: IHC with the monoclonal antibody used in this study is a suitable additional method but no alternative to the direct detection of M. hyopneumoniae by nPCR. Clinical relevance: In the case of the detection of M. hyopneumoniae in BALF by nPCR, typical pathological lesions can be expected with a higher probability than in nPCR-negative piglets.

Zusammenfassung:

Gegenstand und Ziel: Die meisten der wenigen Studien zum Nachweis von M.-hyo pneumoniae-(M.-hyo.-)Infektionen mittels PCR bei jungen Ferkeln erlauben keine Unterscheidung zwischen latenten Infektionen und solchen mit typischen pathomorphologischen Veränderungen. Die Studie bewertet das Vorkommen von M.-hyo.-Infektionen bei Ferkeln anhand der Auswertung von makroskopischen, histologischen, molekularbiologischen und immunhistochemischen Be funden. Die Beurteilung der Eignung einer Immunhistochemie (IHC) als Alternative oder Ergänzung zu anderen Tests war ein weiteres Ziel. Das Studiendesign lässt keine Rückschlüsse auf die Prävalenz von M.-hyo.-Infektionen bei jungen Ferkeln zu. Material und Methoden: 96 erkrankte Ferkel aus acht präselektierten Herden mit erhöhtem Risiko für M.-hyo.- Infektionen wurden zur Sektion ausgewählt. Die Lungen wurden makroskopisch und histologisch untersucht. Ferner erfolgte eine Untersuchung von bronchoalveolärer Lavageflüssigkeit (BALF) und Lungengewebe mittels nested PCR (nPCR) und IHC zum Nachweis von M. hyo. Ergebnisse: Bei 16 der 33 nPCR-positiven Ferkel ließ sich immun - histochemisch Antigen nachweisen, während bei 63 nPCR-negativen Ferkeln auch die IHC negativ verlief (Kappa-Index = 0,55). Von den 17 nPCR-positiven, aber IHC-negativen Ferkeln stammten 12 Tiere aus derselben Herde. Dies führte zu der Annahme, dass nicht alle Feldstämme mit dem verwendeten monoklonalen Antikörper nachweisbar waren. Der Fisher‘s Exakt-Test zeigte einen signifikanten Zusammenhang (p < 0,0001) zwischen dem Nachweis von M. hyo. mittels nPCR in BALF oder mittels IHC in Lungengewebe und dem Vorkommen von typischen pathologischen Veränderungen. Schlussfolgerung: Die IHC mit dem in dieser Studie eingesetzten monoklonalen Antikörper ist ein geeignetes ergänzendes Untersuchungsverfahren, aber keine Alternative zu dem direkten Nachweis von M. hyo. mittels nPCR. Klinische Relevanz: Bei Ferkeln mit Nachweis von M. hyo. in BALF mittels nPCR können typische pathologische Veränderungen mit einer höheren Wahrscheinlichkeit erwartet werden als bei Ferkeln mit negativem nPCR-Befund.

Key words:

Diagnostic - enzootic pneumonia - monoclonal antibody - piglets

Schlüsselwörter:

Diagnostik - enzootische Pneumonie - monoklonaler Antikörper - Ferkel
 

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