Diversity of Clostridium perfringens toxin-genotypes from dairy farms

BMC Microbiology

Table 2 Oligonucleotide primers for C. perfringens toxin gene detection

Primer	Sequence (5′–3′)	Amplificationproduct size [bp]	Primer concentration [mM]	Toxin	Coding toxin gene
Multiplex PCR (major toxin genes)^a
CPA5L	AGTCTACGCTTGGGATGGAA	900	200	alpha	cpa
CPA5R	TTTCCTGGGTTGTCCATTTC		200
CPBL	TCCTTTCTTGAGGGAGGATAAA	611	400	beta	cpb
CPBR	TGAACCTCCTATTTTGTATCCCA		400
CPIL	AAACGCATTAAAGCTCACACC	293	200	iota	iap
CPIR	CTGCATAACCTGGAATGGCT		200
CPETXL	TGGGAACTTCGATACAAGCA	396	200	epsilon	etx
CPETXR	TTAACTCATCTCCCATAACTGCAC		200
Duplex PCR (minor toxin genes)^a
CPEL	GGGGAACCCTCAGTAGTTTCA	506	400	entero	cpe
CPER	ACCAGCTGGATTTGAGTTTAATG		400
CPB2L	CAAGCAATTGGGGGAGTTTA	200	200	beta-2	cpb2
CPB2R	GCAGAATCAGGATTTTGACCA		200
Singleplex PCR for cpa detection^b
224	AGGAACTCATGCTATGATTGTAACTCAAGG	775	200	alpha	cpa
972inverse	ACCACTAGTTGATATGTAAGCTACTAG		200
PCR for cpb2 variants^c
CPB2ATYPF	ATTATGTTTAGGAATACAGTTA	304	200	beta-2 atypical	cpb2aty
CPB2CONF	CAATTGGGGGAGTTTATCCACAA	741	200	beta-2 consensus	cpb2con
CPB2R	CAATACCCTTCACCAAATACTC		200

^aBaums CG, Schotte U, Amtsberg G, Goethe R. Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Vet Microbiol. 2004;100:11–16
^bSchlapp T, Blaha I, Bauerfeind R, Wieler LH, Schoepe H, Weiss R, Baljer G. Synthesis and evaluation of a nonradioactive gene probe for the detection of Clostridium-perfringens alpha-toxin. Mol Cell Probe. 1995;9:101–109
^cJost BH, Billington SJ, Trinh HT, Bueschel DM, Songer JG. Atypical cpb2 genes, encoding beta2-toxin in Clostridium perfringens isolates of nonporcine origin. Infect Immun. 2005;73(1):652–6

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