Development of a multiplex and real time PCR assay for the specific detection of Arcobacter butzleri and Arcobacter cryaerophilus
| Autor: | Robyn Clemens, Jackie Boerema, Gale Brightwell, David Pulford, Stephen L. W. On, Eilidh Mowat |
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| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Microbiology (medical) Meat Arcobacter cryaerophilus Biology Polymerase Chain Reaction Sensitivity and Specificity Microbiology law.invention law Multiplex polymerase chain reaction TaqMan Animals Multiplex Molecular Biology Polymerase chain reaction Arcobacter Sheep rpoB biology.organism_classification Molecular biology Arcobacter butzleri RNA Ribosomal 23S Cattle Gram-Negative Bacterial Infections |
| Zdroj: | Journal of Microbiological Methods. 68:318-325 |
| ISSN: | 0167-7012 |
| DOI: | 10.1016/j.mimet.2006.09.008 |
| Popis: | A new multiplex PCR and two specific TaqMan assays were developed to target the emerging pathogens A. butzleri and A. cryaerophilus. The assays also included an internal control to verify the presence of bacterial target DNA and amplification integrity. The multiplex assay used a published primer set (CRY1 and CRY2) for detecting A. cryaerophilus DNA (Houf, K., Tutenel, A., De Zutter, L., Van Hoof, J. and Vandamme, P., 2000. Development of a multiplex PCR assay for the simultaneous detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii. FEMS microbiology letters, 193 (1): 89-94.) and a novel A. butzleri primer set designed to target the rpoB/C gene sequences. To improve sample throughput and assay sensitivity a TaqMan assay for each Arcobacter spp. was developed which again utilised the heterogeneity contained in the rpoB/C and 23s rRNA gene sequences. The two TaqMan assays provided >2 log improvement in detection sensitivity for both Arcobacter spp. compared with the multiplex PCR assay and were able to detect |
| Databáze: | OpenAIRE |
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