Design and Construction of a Single-Tube, LATE-PCR, Multiplex Endpoint Assay with Lights-On/Lights-Off Probes for the Detection of Pathogens Associated with Sepsis

Table 3

LATE-PCR 16S sepsis multiplex primer and probe sequences.

Primer	Sequence 5′→3′	Bases
16S limiting primer	CCAGACTCCTACGGGAGGCAGCAGT	25
16S excess primer	GTATTACCGCGGCTGTGGCA	20
Candida limiting Primer APT	ACCATTACCTCATTATTGGAGACGTGATGCTGC	33
Candida excess Primer APT	GCAATTTCTTGATCAGTCATTTTTACACCATCTT	34
Candida limiting Primer G	ATGCCCAACAAGCTATCTCTGGTACTTACATGT	33
Candida excess Primer G	CGGATGTTGCAGGGGAAGTATGTTGACCACCCA	33
L. lactis (control) Limiting Primer	CTAAAATCAGGAACTTCGTTATCTTTAGTAGTCACAACCA	40
L. lactis (control) Excess Primer	TAATCATTATTCCTCAAGAAGAGATACAATCGGTCA	36
Control probe	Cal Org 560-ATAAACCTTTCTTAAAAT-BHQ1	18
Candida APT probe	Cal Red 610-ATGTGATATTGATCCAAATCGTGATTTAATAT-BHQ2	32
Candida G probe	Quasar 670-AAACAAGGATGGTACTAGGATGACCGTT-BHQ-2	28
16S Quasar 1 On	Quasar 670-AAGCGAAAGCCTGATGCAGCCATT-BHQ2	24
16S Quasar 2 On	BHQ2-TAGCCGCGTGTGTGAAGAATA-Quasar 670	21
16S Quasar 3 On	Quasar 670-TTATATGTGTAAGTAACTGTGCACATCAA-BHQ2	29
16S Quasar 4 On	Quasar 670-TTGAAGAAGCACCGGCTAACTCCGAA-BHQ2	27
16S Quasar 1 Off	AAGGGGAATATTGCACAATGGTT-BHQ2	23
16S Quasar 2 Off	BHQ2-TTGGCCTTCGGATTGTAAAGCACTTAA-C3 Carbon Linker	27
16S Quasar 3 Off	TATTAGTAGGGAGGAAGTA-BHQ2	19
16S Quasar 4 Off	TTGACGTTACCCGCAA-BHQ2	16