Multiplex detection of bacterial respiratory pathogens

A composition for the detection of S. pneumoniae and N. meningitidis in a sample, comprising:

• at least one pair of primers capable of hybridizing to the lytA gene of S. pneumoniae, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 1-10, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 1-10;
• at least one pair of primers capable of hybridizing to the cpsA gene of S. pneumoniae, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 16-25, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 16-25;
• at least one pair of primers capable of hybridizing to the sodC gene of N. meningitidis, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 31-39, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 31-39;
• and
• at least one pair of primers capable of hybridizing to the crtA gene of N. meningitidis, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 45-54, or sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 45-54.

A method of detecting S. aureus, H. influenzae, M. catarrhalis, and K. pneumoniae in a sample, comprising:

• contacting said sample with a plurality of pairs of primers, wherein the plurality of pairs of primers comprises:
• at least one pair of primers capable of hybridizing to the nuc gene of S. aureus, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 75-83, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 75-83;
• at least one pair of primers capable of hybridizing to the fucK gene of H. influenzae, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 88-95, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 88-95;
• at least one pair of primers capable of hybridizing to the copB gene of M. catarrhalis, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 101-110, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 101-110;
• and
• at least one pair of primers capable of hybridizing to the gltA gene of K. pneumoniae, wherein each primer in said at least one pair of primers comprises any one of the sequences of SEQ ID NOs:
• 116-124, or a sequence that exhibits at least about 85% identity to any one of the sequences of SEQ ID NOs:
• 116-124;
• generating amplicons of the nuc gene sequence of S. aureus, amplicons of the fucK gene sequence of H. influenzae, amplicons of the gene sequence of the copB gene sequence of M. catarrhalis, amplicons of the gltA gene sequence of K. pneumoniae from said sample, or any combination thereof, if said sample comprises one or more of S. aureus, H. influenzae, M. catarrhalis, and K. pneumoniae;
• and
• determining the presence or amount of one or more amplicons as an indication of the presence of one or more of S. aureus, H. influenzae, M. catarrhalis, and K. pneumoniae in said sample.