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Table 1 Primers and probes for multiplex qPCR

From: Reliable detection of Bacillus anthracis, Francisella tularensis and Yersinia pestis by using multiplex qPCR including internal controls for nucleic acid extraction and amplification

Organism

Target

Oligo function

Oligo name

Sequence 5'-3' a

B. anthracis

sspE

Forward primer

spEpri_f

CGACTGAAACAAATGTACAAGCAGTA

  

Reverse primer

spEpri_r

CGTCTGTTTCAGTTGCAAATTCTG

  

Probe

Tqpro_spE

FAM-TGCTAGCATTCAAAGCACAAATGCTAGTT-BHQ1

 

cya

Forward primer

cyapri_f

AGGTAGATTTATAGAAAAAAACATTACGGG

  

Reverse primer

cyapri_r

GCTGACGTAGGGATGGTATT

  

Probe

Tqpro_cya

JOE- CCACTCAATATAAGCTTTATTACCAGGAGC-BHQ1

 

capB

Forward primer

caBpri2_f

AGCAAATGTTGGAGTGATTGTAAATG

  

Reverse primer

caBpri2_r

AAAGTAATCCAAGTATTCACTTTCAATAG

  

Probe

Tqpro_caB

CFR590- AGGTCCCATAACATCCATATGATCTTCTAA-BHQ2

F. tularensis

fopA

Forward primer

foApri_f

GCGCTTTGACTAACAAGGACA

  

Reverse primer

foApri_r

CCAGCACCTGATGGAGAGTT

  

Probe

Tqpro_foA

FAM- TGGCCAGTGGTACTTAGGTGTAGATGCTA-BHQ1

 

ISFtu2

Forward primer

isfpri2_f

CAAGCAATTGGTAGATCAGTTGG

  

Reverse primer

isfpri2_r

GACAACAATATTTCTATTGGATTACCTAAA

  

Probe

Tqpro_isf

JOE- ACCACTAAAATCCATGCTATGACTGATG-BHQ1

 

pdpD

Forward primer

pdDpri_f

TCAATGGCTCAGAGACATCAATTAAAAGAA

  

Reverse primer

pdDpri_r

CACAGCTCCAAGAGTACTATTTCC

  

Probe

Tqpro_pdD

CFR590- ACCAAATCAAAATCCTGCTGAGCAGA-BHQ2

Y. pestis

ypo393

Forward primer

yp93pri_f

AGATAGTGTGACTGGTCTTGTTTCA

  

Reverse primer

yp93pri_r

AGATGCAGATTGTATTGTAAACAATGAC

  

Probe

Tqpro_yp93

FAM- ACTTCCTGATATATTGGAAATCTTCTTCTC-BHQ1

 

caf1

Forward primer

cafpri_f

CCAGCCCGCATCACT

  

Reverse primer

cafpri_r

ATCTGTAAAGTTAACAGATGTGCTAGT

  

Probe

Tqpro_caf

JOE- AGCGTACCAACAAGTAATTCTGTATCGATG-BHQ1

 

pla

Forward primer

plapri_f

ATGAGAGATCTTACTTTCCGTGAGAA

  

Reverse primer

plapri_r

GACTTTGGCATTAGGTGTGACATA

  

Probe

Tqpro_pla

CFR590- TCCGGCTCACGTTATTATGGTACCG-BHQ2

B. thuringiensis

cry1

Forward primer

crypri_f

GCAACTATGAGTAGTGGGAGTAATTTAC

  

Reverse primer

crypri_r

TTCATTGCCTGAATTGAAGACATGAG

  

Probe

Tqpro_cry

Cy5-ACGTAAATACACT-BHQ2- TGATCCATTTGAAAAG-P

  1. a CFR590 = CALFluor Red 590, BHQ = Black Hole quencher, P = phosporylation