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Evaluation of 5' nuclease assay for detection of Actinobacillus pleuropneumoniae.

ABSTRACT: Sequence detection by the 5' nuclease TaqMan assay uses online detection of internal fluorogenic probes in closed PCR tubes. Primers and probe were chosen from a part of the omlA gene common to all serotypes of Actinobacillus pleuropneumoniae, which gave an amplicon of 92 bp. The test was evaluated with 73 lung isolates and 120 tonsil isolates of A. pleuropneumoniae as well as with a collection of reference strains. By using a C(t) value (cycle number in which the fluorescence exceeds the threshold defined by the software) of 30 as the cutoff limit, the 5' nuclease assay represents a test with 100% sensitivity and 100% specificity. A high degree of reproducibility of the test was demonstrated. If samples with C(t) values of

SUBMITTER: Angen O

PROVIDER: S-EPMC87711 | biostudies-literature | 2001 Jan

REPOSITORIES: biostudies-literature

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Evaluation of 5' nuclease assay for detection of Actinobacillus pleuropneumoniae.

Angen O O Jensen J J Lavritsen D T DT

Journal of clinical microbiology 20010101 1

Sequence detection by the 5' nuclease TaqMan assay uses online detection of internal fluorogenic probes in closed PCR tubes. Primers and probe were chosen from a part of the omlA gene common to all serotypes of Actinobacillus pleuropneumoniae, which gave an amplicon of 92 bp. The test was evaluated with 73 lung isolates and 120 tonsil isolates of A. pleuropneumoniae as well as with a collection of reference strains. By using a C(t) value (cycle number in which the fluorescence exceeds the thresh ...[more]

PMID: 11136780

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