O-serogroups, virulence genes, antimicrobial susceptibility, and MLST genotypes of Shiga toxin-producing Escherichia coli from swine and cattle in Central China

Peng, Zhong; Liang, Wan; Hu, Zizhe; Li, Xiaosong; Guo, Rui; Hua, Lin; Tang, Xibiao; Tan, Chen; Chen, Huanchun; Wang, Xiangru; Wu, Bin

doi:10.1186/s12917-019-2177-1

Table 2 Primers used in this study

From: O-serogroups, virulence genes, antimicrobial susceptibility, and MLST genotypes of Shiga toxin-producing Escherichia coli from swine and cattle in Central China

Primers	Sequences (5′-3′)	Annealing Temp. (°C)	Product size (bp)	Function	References
Bacterial identification and virulence genotyping
stx₁-F	ACACTGGATGATCTCAGTGG	60	614	Amplifying stx1	Botteldoorn et al., 2003^b
stx₁-R	CTGAATCCCCCTCCATTATG	60	614	Amplifying stx1	Botteldoorn et al., 2003^b
stx₂-F	GGCACTGTCTGAAACTGCTCC	64	255	Amplifying stx2	Leung et al., 2001^d
stx₂-R	TCGCCAGTTATCTGACATTCTG	64	255	Amplifying stx2	Leung et al., 2001^d
16S-F	ATGGCTCAGATTGAACGC	50	1505	Amplifying 16 SrRNA	REN et al., 2012^g
16S-R	CAGGTTCCCCTACGGTTA	50	1505	Amplifying 16 SrRNA	REN et al., 2012^g
eae-F	GTGGCGAATACTGGCGAGACT	64	890	Amplifying eae	Nielsen et al., 2003^e
eae-R	CCCCATTCTTTTTCACCGTCG	64	890	Amplifying eae	Nielsen et al., 2003^e
ehxA-F	GCATCATCAAGCGTACGTTCC	60	534	Amplifying ehxA	Bandyopadhyay et al., 2011^a
ehxA-R	AATGAGCCAAGCTGGTTAAGCT	60	534	Amplifying ehxA	Bandyopadhyay et al., 2011^a
saa-F	CCTCACATCTTCTGCAAATACC	60	1688	Amplifying saa	Paton et al., 2001^f
saa-R	GTTGTCGTTCATATTTTACCATCCAATGGACATG	60	1688	Amplifying saa	Paton et al., 2001^f
MLST genotyping
Adk-F1	TCATCATCTGCACTTTCCGC	54	583	Amplifying adk	Ding et al., 2012^c
Adk-R1	CCAGATCAGCGCGAACTTCA	54	583	Amplifying adk
FumC-F1	TCACAGGTCGCCAGCGCTTC	54	806	Amplifying fumC
FumC-R1	GTACGCAGCGAAAAAGATTC	54	806	Amplifying fumC
GyrB-F1	TCGGCGACACGGATGACGGC	60	911	Amplifying gyrB
GyrB-R1	ATCAGGCCTTCACGCGCATC	60	911	Amplifying gyrB
Icd-F1	ATGGAAAGTAAAGTAGTTGTT CCGGCACA	54	878	Amplifying icd
Icd-R1	GGACGCAGCAGGATCTGTT	54	878	Amplifying icd
Mdh-F1	ATGAAAGTCGCAGTCCTCGGC GCTGCTGGCGG	60	932	Amplifying mdh
Mdh-R1	TTAACGAACTCCTGCCCCAGAGCGATATCTTTCTT	60	932	Amplifying mdh
PurA-F1	TCGGTAACGGTGTTGTGCTG	54	816	Amplifying purA
PurA-R1	CATACGGTAAGCCACGCA GA	54	816	Amplifying purA
RecA-F1	CGCATTCGCTTTACCCTGACC	58	780	Amplifying recA
RecA-R1	TCGTCGAAATCTACGGACCGGA	58	780	Amplifying recA
Adk-F2	TCATCATCTGCACTTTCCGC	–	–	adk Sequencing
Adk-R2	CCAGATCAGCGCGAACTTCA	–	–	adk Sequencing
FumC-F2	TCACAGGTCGCCAGCGCTTC	–	–	fumC Sequencing
FumC-R2	TCCCGGCAGATAAGCTGTGG	–	–	fumC Sequencing
GyrB-F2	TCGGCGACACGGATGACGGC	–	–	gyrB Sequencing
GyrB-R2	GTCCATGTAGGCGTTCAGGG	–	–	gyrB Sequencing
Icd-F2	ATGGAAAGTAAAGTAGTTGTTCCGGCACA	–	–	icd Sequencing
Icd-R2	GGACGCAGCAGGATCTGTT	–	–	icd Sequencing
Mdh-F2	AGCGCGTTCTGTTCAAATGC	–	–	mdh Sequencing
Mdh-R2	CAGGTTCAGAACTCTCTCTGT	–	–	mdh Sequencing
PurA-F2	CGCGCTGATGAAAGAGATGA	–	–	purA Sequencing
PurA-R2	CATACGGTAAGCCACGCAGA	–	–	purA Sequencing
RecA-F2	ACCTTTGTAGCTGTACCACG	–	–	recA Sequencing
RecA-R2	TCGTCGAAATCTACGGACCGGA	–	–	recA Sequencing

^aBandyopadhyay S, Mahanti A, Samanta I, Dutta TK, Ghosh MK, Bera AK, Bandyopadhyay S, Bhattacharya D. Virulence repertoire of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic Escherichia coli (ETEC) from diarrhoeic lambs of Arunachal Pradesh, India. Trop Anim Health Prod. 2011;43(3):705-10
^bBotteldoorn N, Heyndrickx M, Rijpens N, Herman L. Detection and characterization of verotoxigenic Escherichia coli by a VTEC/EHEC multiplex PCR in porcine faeces and pig carcass swabs. Res Microbiol. 2003;154(2):97-104
^cDing Y, Tang X, Lu P, Wu B, Xu Z, Liu W, Zhang R, Bei W, Chen H, Tan C. Clonal analysis and virulent traits of pathogenic extraintestinal Escherichia coli isolates from swine in China. BMC Vet Res. 2012;8:140
^dLeung PH, Yam WC, Ng WW, Peiris JS. The prevalence and characterization of verotoxin-producing Escherichia coli isolated from cattle and pigs in an abattoir in Hong Kong. Epidemiol Infect. 2001;126(2):173-9
^eNielsen EM, Andersen MT. Detection and characterization of verocytotoxin-producing Escherichia coli by automated 5' nuclease PCR assay. J Clin Microbiol. 2003;41(7):2884-93
^fPaton AW, Srimanote P, Woodrow MC, Paton JC. Characterization of Saa, a novel autoagglutinating adhesin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun. 2001;69(11):6999-7009
^gREN L, YU X, SONG D, ZHEN K, QIN Y, WANG Y. Isolation, identification and phylogenetic analysis of E. coli from Yaks. China Animal Husbandry & Veterinary Medicine. 2012;39(1): 168-171

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