Table 2 Targets, primer sequences and final concentrations for each of the species-specific quintuplex HRM assays
Brucella species | Forward primer sequence | Reverse primer sequence | Final concentration of primer pair in quintuplex reaction (μM) | Product size (bp) | Gene target (based on B. abortus 9-941) |
|---|---|---|---|---|---|
B. abortus | 5′-GCCCCTCCTTCTTGTAATCA-3′ | 5′-ACCATGAAGAAAGCGCGTAT-3′ | 1.25 μM | 75 | BruAb1_0395Hypothetical protein |
B. melitensis | 5′-ACAAGCTGACGAAGGACCAT-3′ | 5′-CCCGTATAGGAGTGGATCGT- 3′ | 0.5 μM | 135 | BruAb1_1713 glyceraldehyde-3-phosphate dehydrogenase |
B. ovis | 5′-CCGGTCAAGTTCAATCACG-3′ | 5′-GCTGGAAATGCTCTATTACTC-3′ | 1 μM | 66 | BruAb1_1179 Hypothetical protein |
B. suis | 5′-CTGGCGGAAAAGGATTTGAT-3′ | 5′-AATCACGACAAACCACAGCA-3′ | 1.125 μM | 89 | BruAb1_1338 Sugar ABC transporter, permeaseprotein |
B. canis | 5′-CCCCCGTCAATTCCTGCCGAA-3′ | 5′-CCCCCGTGGCCTGGTCGAGAT-3′ | 0.25 μM | 79 | BruAb2_1115 Transcriptional regulator (GntR Family) |