. 2020 Jul 10;52(6):3173–3185. doi: 10.1007/s11250-020-02343-1

Table 3.

Components and amplification PCR conditions utilized for detecting genes encoding virulence factors

Genes encoding virulence factors	PCR components and volume (μl)	PCR conditions	References
eae, hlyA, stx₁, and astA	5 μL Master Mix 5 μL DNA template 0.5 μL of each F&R primer (with total 4 μL) 11 μL PCR-grade water	First condition 1 cycle [94 °C, 5 min], 35 cycles [94 °C, 30 s/62 °C, 30 s/72 °C, 1 min], and 1 cycle [72 °C, 5 min]	Chandra et al. (2013)
stx₂	5 μL Master Mix 5 μL DNA template 13 μL PCR grade water 1 μL of F&R primers (with total 2 μL)	Second condition 1 cycle [95 °C, 3 min], 35 cycles [95 °C, 20 s/58 °C, 40 s/72 °C, 90 s], and 1 cycle [72 °C, 5 min]	Gannon et al. (1992); Kamel et al. (2015)
lt, st, and f5	5 μL Master Mix 5 μL DNA template 12 μL PCR-grade water 0.5 μL of each F&R primers (with total 3 μL)	3rd condition 1 cycle [95 °C, 5 min], 40 cycles [95 °C, 45 s/50 °C, 1 min/72 °C, 1 min], and 1 cycle [72 °C, 7 min]	Aranda et al. (2004)

Genes encoding virulence factors

PCR components and volume (μl)

PCR conditions

References

eae, hlyA, stx₁, and astA

5 μL Master Mix

5 μL DNA template

0.5 μL of each F&R primer (with total 4 μL)

11 μL PCR-grade water

First condition

1 cycle [94 °C, 5 min], 35 cycles [94 °C, 30 s/62 °C, 30 s/72 °C, 1 min], and 1 cycle [72 °C, 5 min]

Chandra et al. (2013)

stx₂

5 μL Master Mix

5 μL DNA template

13 μL PCR grade water

1 μL of F&R primers (with total 2 μL)

Second condition

1 cycle [95 °C, 3 min], 35 cycles [95 °C, 20 s/58 °C, 40 s/72 °C, 90 s], and 1 cycle [72 °C, 5 min]

Gannon et al. (1992); Kamel et al. (2015)

lt, st, and f5

5 μL Master Mix

5 μL DNA template

12 μL PCR-grade water

0.5 μL of each F&R primers (with total 3 μL)

3rd condition

1 cycle [95 °C, 5 min], 40 cycles [95 °C, 45 s/50 °C, 1 min/72 °C, 1 min], and 1 cycle [72 °C, 7 min]

Aranda et al. (2004)