TABLE 1

Primers and probes utilized for specific PCR detection of pathogens

TargetPrimers and probeReference(s)
Haemophilus influenzae, 16S rRNA geneForward primer, 5′-CGTATTATCGGAAGATGAAAGTGC-3′; reverse primer, 5′-CTACGCATTTCACCGCTACAC-3′; hydrolysis probe, 5′-FAM-CGTATTACCGCGGCTGCTGGCAC-3′aPrimers, 16, 24, and 32; probe, 17
Streptococcus pneumoniae, oralis, mitis and infantis, 16S rRNA geneForward primer, 5′-AAGGTGCACTTGCATCACTACC-3′; reverse primer and probe as in the assay for H. influenzae
Alloiococcus otitidis, 16S rRNA geneForward primer, 5′-GGGGAAGAACACGGATAGGA-3′; reverse primer and probe as in the assay for H. influenzae
Moraxella catarrhalis, 16S rRNA geneForward primer, 5′-CCCATAAGCCCTGACGTTAC-3′; reverse primer and probe as in the assay for H. influenzae
Streptococcus pneumoniae, lytA geneForward primer, 5′-ACGCAATCTAGCAGATGAAGCA-3′; reverse primer, 5′-TCGTGCGTTTTAATTCCAGCT-3′; hydrolysis probe, 5′-FAM-TGCCGAAAACGCTTGATACAGGGAG-3′21
Pseudomonas aeruginosa, oprL geneForward primer, 5′-GGGTTCATTAGGAGTTACATGA-3′; reverse primer, 5′-GGGCATAACGACTTCTTACTTC-3′; hydrolysis probe, 5′-FAM-TTCTGAGCCCAGGACTGCTCGTCGTGGCCG-3′Primers repositioned from those reported in reference 18
Staphylococcus aureus, nuc geneForward primer, 5′-CATCCTAAAAAAGGTGTAGAGA-3′; reverse primer, 5′-TTCAATTTTMTTTGCATTTTCTACCA-3′; hydrolysis probe, 5′-FAM-TTTTCGTAAATGCACTTGCTTCAGGACCA-3′19
  • a The reverse primer and the hydrolysis probe are common to the first four assays. FAM, 6-carboxyfluorescein.