Nested-PCR for detection of Brucella in milk
To detect Brucella inflection in milk, a nested-PCR assay was developed by designing outward-directed and inward-directed primers for omp25, omp31 and 16S rRNA genes of Brucella abortus, and optimizing the extraction of Brucella genomic DNA. The results show that two rounds of amplification with the...
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| Główni autorzy: | , , , , |
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| Format: | Artykuł |
| Język: | angielski |
| Wydane: |
Zhejiang University Press
2008-03-01
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| Seria: | 浙江大学学报. 农业与生命科学版 |
| Hasła przedmiotowe: | |
| Dostęp online: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.02.008 |
| Etykiety: |
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| Streszczenie: | To detect Brucella inflection in milk, a nested-PCR assay was developed by designing outward-directed and inward-directed primers for omp25, omp31 and 16S rRNA genes of Brucella abortus, and optimizing the extraction of Brucella genomic DNA. The results show that two rounds of amplification with the two pairs of synthesized primers F4/R2 and F2D/R1U can effectively detect the specific DNA fragment of 16S rRNA from the Brucella contaminated milk, of which the detection limit was as low as 3.3 CFU-mL<sup>-1</sup>. This PCR assay showed a high specificity for Brucella with no cross-reaction of the related bacteria DNA, including Escherichia coli, Hemolytic streptococci, Salmonella, Klebsiella, Staphylococcus aureus, Pseudomonas aeruginosa, and Helicobacter pylori. The developed nested-PCR assay has 100% identity to Brucella isolation. Therefore, it is suggested that the nested-PCR assay for Brucella with high specificity and sensitivity is potential to detect Brucella infection of milk in laboratory. |
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| ISSN: | 1008-9209 2097-5155 |