Table 2.
Oligonucleotide primers used for multiplex PCRs to detect 16 genes putatively linked with virulence of E. rhusiopathiae
| Gene / locus tag | Gene product / predicted function [reference] | Primer sequence (5′ – 3′) (forward/reverse)a | GenBank Acc. No | Localization within gene | Size (bp) | Gene pre-valence (%) |
|---|---|---|---|---|---|---|
| ERH_1356 | ABC transporter metal-binding protein / adhesion of host cells [45] | CATGAAGGGTAACACCTTGG/ GGGCGATAAAGTTGCGGTAGAA | NC_015601.1 | 579 – 787 | 209 | 100 |
| intI-like | Internalin / invasion of epithelial cells [18,45] | ACAGTTTCGGATACTTCCGG/ ACCCTCGTCATATTTACCAGC | AP012027.1 | 386 – 714 | 329 | 85.5 |
| rspB | Rhusiopathiae surface protein /biofilm formation [41] | ATCTTTACCCAATTCGACGT/ ATGAACCCAGTCCAAGATTGG | AB052682.1 | 6882 – 7287 | 406 | 100 |
| rspA | Rhusiopathiae surface protein /biofilm formation [41] | ATCGACTGGTATTCAGTTGG/ ATCACGAGACATACCGCCAA | AB052682.1 | 597 – 1131 | 535 | 100 |
| cap locus | Capsule / resistance to phagocytosis, intracellular survival [40] | TATCTTTGTAGCGTAGTTGG/ CAATAAAAGGAAATACCAGTGC | D64177 | 1333 – 1987 | 645 | 100 |
| algI | Alginate-O-acetyltransferase / resistance to phagocytosis [45] | AGTTATCTTGGACTTGGTCC/ AGATAAGTGCGCATTGATCC | AP012027.1 | 121 – 887 | 767 | 97.6 |
| ewlA | Lipoprotein / adhesion to host cells [45] | TAATATTAGATAGCGAGGAAT/ AAGAAAAGGGAGTGTGAATAT | U52850.1 | 226 –1185 | 960 | 100 |
| nanH | Neuraminidase / spreading factor, nutritive [44] | ATGAAGCGCTTACATTTGAAT/ TACATAAGGTTGACCAAAGTC | AB019122 | 295 –1401 | 1.107 | 100 |
| sub | S8-subtilisin / peptidase [45] | AAGCCTGAGATATCTGCACC/ TTGTACAATTGGATGAGCCG | AP012027.1 | 1360 –1585 | 226 | 100 |
| sodA | Superoxide dismutase / antioxidans [45,70] | AGAAGACATCCGCACAGCAGT/ GCATGTTCCCAAACATCAAGA | AP012027.1 | 195 – 509 | 315 | 100 |
| mviN1 | Integral membrane protein / cell adhesion, transport protein [45] | AAATCATGCTTGTAATGGCGG/ ATTCGACGTTAAAACAACCGC | AP012027.1 | 565 –1032 | 378 | 100 |
| hep | Heparinase / inactivation of heparin [45] | ATGGAAGTACCGATCTCACT/ TCATTGTAGCAACATGGCTTC | AP012027.1 | 997 –1480 | 484 | 100 |
| hlyIII | Hemolysin / lytic activity on red blood cells [42] | TACGATTGCGACAAAGTGTGCG/ ATGGAAACATAGGGAAGGCTG | AP012027.1 | 16 – 559 | 544 | 100 |
| fbpA | Fibronectin-binding protein / adhesion [41] | ATCTCGCCGCTTTTAGAACG/ GCGTCTTCAACTGTTGCTTG | AP012027.1 | 565 –1166 | 602 | 100 |
| hlyA | Hyaluronidase / spreading factor [2] | AGGATCACTTACCGCTATGG/ CAGCACTCAGCATGTTCTC | AP012027.1 | 598 –1538 | 941 | 100 |
| dnaB | Membrane-, attachment protein / proliferation, adhesion [45] | AATAGCCCCTGATCAAATGG/ CTCTCCTTTACTTAACATCCC | ACLK02000002.1 | 39 –1155 | 1.117 | 99.4 |
Primers were designed in this study except for the primers to amplify the cap locus [46].
aIn case Ogawa et al. [45] is given as a reference, the predicted function of the gene product has been deduced from genome annotation data of strain Fujisawa. Here, in vitro or in vivo studies would be mandatory to verify the linkage of the genes with the pathogenesis of Erysipelas in different hosts.