TABLE 2.

Primers and probes used in mPCR/RLB for detection of 13 toxin genes

Primer/probeaTargetTm °CbGenBank accession no.Primer/probe sequence (5′-3′)cReferencesd
nucSbnuc65.68V01281511GCG ATT GAT GGT GAT ACG GTT5317
nucApnuc61.36V01281558CAT TGG TTG ACC TTT GTA CAT TAA 535This study
nucSpnuc61.06V01281745GAT GGA AAA ATG GTA AAC GAA G766This study
nucAbnuc69.12V01281789AGC CAA GCC TTG ACG AAC TAA AGC7667
seaSbsea64.05M18970487CCT TTG GAA ACG GTT AAA ACG5073
seaSpsea68.83M18970531GGA GTT GGA TCT TCA AGC AAG ACG 5543
seaApsea63.87M18970613TCT GAA CCT TCC CAT CAA AAA C5923
seaAbsea62.91M18970691TTGA ATA CTG TCC TTG AGC ACC67043
sebSbseb64.82M11118634TCG CAT CAA ACT GAC AAA CG6533
sebSpseb61.1M11118662GTAT GTA TGG TGG TGT AAC TGA GC68543
sebApseb60.06M11118831CA CCA AAT AGT GAC GAG TTA GG81043
sebAbseb60.4M11118924CAT GTC ATA CCA AAA GCT ATT CTC9013
secSbsec62.5X05815664G CTC AAG AAC TAG ACA TAA AAG CTA GG6903
secSpsec63.13X05815772AAC GG(/a)C AAT ACT TTT TGG TAT GAT7953
secApsec61.4X05815885CTT CAC A(/t)CT TTT AGA ATC AAC CG86343
secAbsec60.4X05815935TCA AAA TCG GAT TAA CAT TAT CC9133
sedSbsed60.2M28521332CTA GTT TGG TAA TAT CTC CTT TAA ACG3583
sedSpsed64.91M28521360TAA AGC CAA TGA AAA CAT TGA TTC A3843
sedApsed60.85M28521491CTT TTA TTT TCT CCT ATT ATT GG ATTTTT46330
sedAbsed61.9M28521653CAA TTA ATG CTA TAT CTT ATA GGG TAA ACA TC6223
seeSbsee63.41M21319424C GAT TGA CCG AAG AAA AAA AAG44530
seeSpsee60.2M21319479CTA CAG TAC CTA TAG ATA AAG TTA AAA CAA GC5103
seeApsee66.87M21319613TTT GCA CCT TAC CGC CAA AG5943
seeAbsee60.38M21319659TAA CTT ACC GTG GAC CCT TC6403
segSbseg66.14AF064773229CAA CCC/T GAT CCT AAA TTA GAC GAA C2532
segSpseg63.09AF064773285GGG AAC TAT GGG T(/a)AA TGT AAT GAA TC3102
segApseg62.61AF064773338CTT CCT TCA ACA GGT GGA GAC3182
segAbseg62.91AF064773485/401GGA ACG CCA AAA ATG TCT ACT T464/37935
sehSbseh60.86U11702407TTA GAA ATC AAG GTG ATA GTG GC 4292
sehSpseh61.25U11702454ACT GCT GAT TTA GCT CAG AAG TTT A 4782
sehApseh60.1U11702575AGT GTT GTA CCT CCA TAT AGA C ATTC55035
sehAbseh60.47U11702641TTT TGA ATA CCA TCT ACC CAA AC6192
seiSbsei63.01AY158703396G GCC ACT TTA TCA GGA CAA TAC TT4192
seiSpsei61.91AY158703656A CA C(a)TG GTA AAG GC(t)A AAG AAT ATG6792
seiApsei62.26AY158703726AAA ACT TAC AGG CAG TCC ATC TC7042
seiAbsei58.23AY158703818AAT TAT CAT TAG TTA CTA TCT ACA TAT GAT ATT TC78435
etaSbeta61.39M17347374CTA GTG CAT TTG TTA TTC AAG ACG3973
etaSpeta69.51M17347414CCA TGC AAA AGC AGA AGT TTC AGC 4373
etaApeta60.67M17347492TGC A(/g)TT GAC ACC ATA GTA CTT ATT C468This study
etaAbeta62.72M17347794AAT GCT AAA TCA ACA CCT GC AC77330
etbSbetb61.26M17348190TAC CAC CTA ATA CCC TAA TAA TCC AA2153
etbSpetb61.37M17348286GAG ACA GTG CAT TAA ATG AAT AAC TTT3123
etbApetb62.41M17348539GAT TTC TTC TGC GCT GTA TTC TT517This study
etbAbetb61.16M17348609C ATT ATC CGT AAT GTG TGT ATAAA GC58443
etdSbetd61.75AB0574215963GCT CGG ATA CCC TTA TAA CTT TTC5986This study
etdSpetd62.2AB0574216055CTG AGT CGG GAA ATT CTG G607343
etdApetd61.47AB0574216120CAA CAT GAA TAC CA0A CTA ACT CTC C6096This study
etdAbetd61.88AB0574216259CAT TAC TAA TGA GAC TGT AAT TCA GCT CT6231This study
tsstSbtsst-165.22J02615348AAG CCA ACA TAC TAG CGA AGG AAC3713
tsstSptsst-160.5J02615394GGC GTT ACA AAT ACT GAA AAA TTA C41830
tsstAptsst-164.36J02615495ATC GAA CTT TGG CCC(/a) ATA CTT T4743
tsstAbtsst-161.03J02615556GTA TTT GAG TTA GCT GAT GAC GAA53343
pvlSbpvl65.29X727002651TTT TAG GCT CAA GAC AAA GCA AC2673This study
pvlAppvl65.3X727002731TAC CTC TGG ATA ACA CTG GCA TTT T270711
pvlSppvl61.76X727002733CTT CAA TCC AGA ATT TAT TGG TGT 275611
pvlAbpvl65.8X727002783TTT GCA GCG TTT TGT TTT CG276411
  • a S, sense; A, antisense; b, biotin labeled (all the primers were biotin labeled at the 5′ end); p, probe (all the probes were 5′ end C6 amine labeled).

  • b Tm values were provided by the primer synthesizer (Sigma-Aldrich).

  • c Boldface numbers represent the numbered base positions at which primer/probe sequences start and finish (starting at point “1” of the corresponding GenBank sequence). Underlined portions indicate modifications of published primer/probe sequences. The bases in parenthesis represent sequences with polymorphisms compared with GenBank sequences or our own sequencing results (for five probes with heterogeneous hybridization).

  • d Primers and probes were used as previously published (some with modification) except, as indicated, those designed for this study.