Prevalence, identification of virulence factors, O-serogroups and antibiotic resistance properties of Shiga-toxin producing Escherichia coli strains isolated from raw milk and traditional dairy products

Reza Ranjbar; Farhad Safarpoor Dehkordi; Mohammad Hossein Sakhaei Shahreza; Ebrahim Rahimi

doi:10.1186/s13756-018-0345-x

Antimicrobial Resistance & Infection Control

Table 1 Primers and PCR conditions used for characterization of virulence genes, O-serogroups and antibiotic resistance genes in the STEC strains isolated from raw milk and traditional dairy product samples

From: Prevalence, identification of virulence factors, O-serogroups and antibiotic resistance properties of Shiga-toxin producing Escherichia coli strains isolated from raw milk and traditional dairy products

Target gene	Primer sequence (5′-3′)	PCR product (bp)	PCR programs	PCR Volume (50 μL)
O157	F: CGGACATCCATGTGATATGG R: TTGCCTATGTACAGCTAATCC	259	1 cycle: 95 ^0C ------------ 3 min.	5 μL PCR buffer 10X 2 mM Mgcl₂
O145	F: CCATCAACAGATTTAGGAGTG R: TTTCTACCGCGAATCTATC	609	30 cycle: 95 ^0C ------------ 20 s 58 ^0C ------------ 40 s 72 ^0C ------------ 30 s	150 μM dNTP (Fermentas) 0.75 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas)
O103	F: TTGGAGCGTTAACTGGACCT R: GCTCCCGAGCACGTATAAG	321
O26	F: CAGAATGGTTATGCTACTGT R: CTTACATTTGTTTTCGGCATC	423
O111	F: TAGAGAAATTATCAAGTTAGTTCC R: ATAGTTATGAACATCTTGTTTAGC	406	1 cycle: 72 ^0C ------------ 8 min	3 μL DNA template
O91	F: GCTGACCTTCATGATCTGTTGA R: TAATTTAACCCGTAGAATCGCTGC	291	1 cycle: 94 ^0C ------------ 6 min. 34 cycle: 95 ^0C ------------ 50 s 58 ^0C ------------ 70 s 72 ^0C ------------ 55 s 1 cycle: 72 ^0C ------------ 10 min	5 μL PCR buffer 10X 2 mM Mgcl₂ 150 μM dNTP (Fermentas) 0.75 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 3 μL DNA template
O128	F: GCTTTCTGCCGATATTTGGC R: CCGACGGACTGATGCCGGTGATT	289
O121	F: TGGCTAGTGGCATTCTGATG R: TGATACTTTAGCCGCCCTTG	322
O113	F: GGGTTAGATGGAGCGCTATTGAGA R: AGGTCACCCTCTGAATTATGGCAG	771
O45	F: CCGGGTTTCGATTTGTGAAGGTTG R: CACAACAGCCACTACTAGGCAGAA	527
stx1	F: AAATCGCCATTCGTTGACTACTTCT R: TGCCATTCTGGCAACTCGCGATGCA	366	1 cycle: 95 ^0C ------------ 3 min. 34 cycle: 94 ^0C ------------ 60 s 56 ^0C ------------ 45 s 72 ^0C ------------ 60 s 1 cycle: 72 ^0C ------------ 10 min	5 μL PCR buffer 10X 2 mM Mgcl₂ 150 μM dNTP (Fermentas) 0.75 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 3 μL DNA template
stx2	F: CGATCGTCACTCACTGGTTTCATCA R: GGATATTCTCCCCACTCTGACACC	282
eaeA	F: TGCGGCACAACAGGCGGCGA R: CGGTCGCCGCACCAGGATTC	629
ehly	F: CAATGCAGATGCAGATACCG R: CAGAGATGTCGTTGCAGCAG	432
aadA1	F: TATCCAGCTAAGCGCGAACT R: ATTTGCCGACTACCTTGGTC	447	1 cycle: 94 ^0C ------------ 8 min. 32 cycle: 95 ^0C ------------ 60 s 55 ^0C ------------ 70 s 72 ^0C ------------ 2 min 1 cycle: 72 ^0C ------------ 8 min	5 μL PCR buffer 10X 2 mM Mgcl₂ 150 μM dNTP (Fermentas) 0.75 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 3 μL DNA template
tetA	F: GGTTCACTCGAACGACGTCA R: CTGTCCGACAAGTTGCATGA	577
tetB	F: CCTCAGCTTCTCAACGCGTG R: GCACCTTGCTGATGACTCTT	634
dfrA1	F: GGAGTGCCAAAGGTGAACAGC R: GAGGCGAAGTCTTGGGTAAAAAC	367
qnr	F: GGGTATGGATATTATTGATAAAG R: CTAATCCGGCAGCACTATTTA	670
aac(3)-IV	F: CTTCAGGATGGCAAGTTGGT R: TCATCTCGTTCTCCGCTCAT	286
sul1	F: TTCGGCATTCTGAATCTCAC R: ATGATCTAACCCTCGGTCTC	822
blaSHV	F: TCGCCTGTGTATTATCTCCC R: CGCAGATAAATCACCACAATG	768
CITM	F: TGGCCAGAACTGACAGGCAAA R: TTTCTCCTGAACGTGGCTGGC	462
cat1	F: AGTTGCTCAATGTACCTATAACC R: TTGTAATTCATTAAGCATTCTGCC	547
cmlA	F: CCGCCACGGTGTTGTTGTTATC R: CACCTTGCCTGCCCATCATTAG	698

Back to article page