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Table 2 Alternate primers used for amplifying and sequencing each of the seven genes for multi locus sequencing typing S. pneumoniae

From: Effectiveness of the standard and an alternative set of Streptococcus pneumoniae multi locus sequence typing primers

Typing gene

Primer sequences

Annealing temperature °C

1 aroE shikimate dehydrogenase

F: 5′-TCCTATTAAGCATTCTATTTCTCCCTTC

55

R: 5′-ACAGGAGAGGATTGGCCATCCATGCCCACACTG

2 gdh glucose-6-phosphate dehydrogenase

F: 5′-ATGGACAAACCAGC(G/A/T/C)AG(C/T)TT

55

R: 5′-GCTTGAGGTCCCAT(G/A)CT(G/A/T/C)CC

2 gki glucose kinase

F: 5′-GGCATTGGAATGGGATCACC

60

R: 5′-TCTCCCGCAGCTGACAC

1,2 recP transketolase

F: 5′-GCCAACTCAGGTCATCCAGG

65

R: 5′-TGCTGTTTCGATAGCAGCATGGATGGCTTCC

3 spi signal peptidase I

F: 5′-GAATTCATTTAAAAATTTCTTAAAAGAGTGG

50

R: 5′-TTAAAATGTTCCGATACGGGTGATTGG

1 xpt xanthine phosphoribosyltransferase

F: 5′-TTAACTTTTAGACTTTAGGAGGTCTTATG

55

R: 5′-CGGCTGCTTGCGAGTGTTTTTCTTGAG

1,3 ddl D-alanine-D-alanine ligase

F: 5′-TAAAATCACGACTAAGCGTGTTCTGG

65

R: 5′-CGCTCGATTAGTTTTGGGTAGCTGATCCC

  1. 1The aroE primers, the recP reverse primer, the xpt primers and the ddl forward primer were designed by the US Centers for Disease Control [12].
  2. 2The recP forward primer, and the gdh and gki primers are the originals developed by Spratt and Enright [11].
  3. 3The spi primers and the ddl reverse primer were designed as part of this study.