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Selected Publication:

Type of publication: Doctoral Thesis
Type of document:

Year: 2004

Authors: Strimitzer, S

Title: Entwicklung und Evaluierung einer Polymerasekettenreaktion (PCR) zum Nachweis von Mycoplasma (Syn. Eperythrozoon) Suis.

Source: Dissertation, Vet. Med. Univ. Wien, pp. 45.

Advisor(s):

Joachim Anja


Abstract:
The study aimed to develop and evaluate a PCR test for detection of Mycoplasma (M.) suis in blood and tissue samples of pigs. Based on the published sequence data of the 16S ribosomal RNA gene of M. suis (GenBank sequence accession no. U88565), 3 sets of species-specific primers were designed for comparative studies on field samples from 187 animals. They were used both separately and combined in nested PCR protocols. For all further investigations the primer combination of the most sensitive nested PCR protocol was selected. With this protocol a 190 base-pair (bp) fragment of the target gene could be amplified to a minimal detection limit of about 250 fg of partially purified, genomic M. suis-DNA. No bands were observed with nucleic acids from M. hyorhinis. When DNA extracted from M. hyopneumoniae and M. hyosynoviae was used as a template, 1 out of 62 runs generated a PCR product of approximately 190 bp length, respectively. Blood smears were stained with acridine orange and evaluated by fluorescence microscopy. The molecular test showed a positive result in 31.6 % of the cases; while a significantly (p<0.001) lower percentage of 12.8 % of the animals was diagnosed positive by blood smear, 4.3 % were positive with either method. Applying the established PCR-based test in further studies, 27.5 % out of 822 pigs were found to be infected with M. suis. With respect to prevalence, parallel investigations of both blood and spleen of 167 animals demonstrated no significant difference (p>0.05) and no correlation (p>0.05) between blood and tissue samples. In the majority of cases (83.8 %) the bacteria were either detectable only in peripheral blood or only in spleen. The presented nested PCR assay proved to be highly sensitive, quick as well as easy to perform and therefore suitable for the routine diagnostic procedure.

Keywords:
pig / eperythrozoonosis / Eperythrozoon suis / Mycoplasma suis / diagnosis / polymerase chain reaction (PCR)


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