Use of a Mycoplasma hyopneumoniae nested polymerase chain reaction test to determine the optimal sampling sites in swine

Kathy L. Kurth, Tsungda Hsu, Eric R. Snook, Eileen L. Thacker, Brad J. Thacker, F. Chris Minion

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

A number of polymerase chain reaction (PCR)-based diagnostic tests have been developed for Mycoplasma hyopneumoniae, including one from this research group. This report presents further development, optimization, and standardization of a nested PCR test. Detection sensitivity was 1 fg of M. hyopneumoniae chromosomal DNA (approximately 1 organism). This exceeded the sensitivity of or compared favorably with other published PCR tests. Polymerase chain reaction primers to porcine β2-microglobulin were included as internal controls for amplifiable chromosomal DNA from porcine samples. To standardize the test, a number of samples from experimentally infected pigs, including nasal, tonsil, tracheobronchial swabs, lung tissue, bronchial alveolar lavage (BAL) fluid, and tracheobronchial brush samples, were examined by PCR. Samples obtained from BAL fluid and tracheobronchial sites were most predictive of infection, whereas nasal swabs and lung tissue were not reliable indicators of experimentally induced infection. In conclusion, the nested PCR developed for this study was found to be a highly sensitive and specific diagnostic tool for M. hyopneumoniae, but the enhanced sensitivity may be unnecessary if the proper sites are sampled.

Original languageEnglish (US)
Pages (from-to)463-469
Number of pages7
JournalJournal of Veterinary Diagnostic Investigation
Volume14
Issue number6
DOIs
StatePublished - Jan 1 2002
Externally publishedYes

Fingerprint

Mycoplasma hyopneumoniae
Swine
polymerase chain reaction
Polymerase Chain Reaction
swine
testing
Bronchoalveolar Lavage Fluid
Nose
sampling
lungs
Lung
tonsils
Palatine Tonsil
DNA
Infection
Routine Diagnostic Tests
standardization
infection
diagnostic techniques
organisms

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Use of a Mycoplasma hyopneumoniae nested polymerase chain reaction test to determine the optimal sampling sites in swine. / Kurth, Kathy L.; Hsu, Tsungda; Snook, Eric R.; Thacker, Eileen L.; Thacker, Brad J.; Minion, F. Chris.

In: Journal of Veterinary Diagnostic Investigation, Vol. 14, No. 6, 01.01.2002, p. 463-469.

Research output: Contribution to journalArticle

Kurth, Kathy L. ; Hsu, Tsungda ; Snook, Eric R. ; Thacker, Eileen L. ; Thacker, Brad J. ; Minion, F. Chris. / Use of a Mycoplasma hyopneumoniae nested polymerase chain reaction test to determine the optimal sampling sites in swine. In: Journal of Veterinary Diagnostic Investigation. 2002 ; Vol. 14, No. 6. pp. 463-469.
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