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Table 3 Comparison of quantitatively detecting Vibrio parahaemolyticus ATCC 27969 in spiked oysters by using the toxR-based LAMP assay in two platforms and PCRa

From: Development of a toxR-based loop-mediated isothermal amplification assay for detecting Vibrio parahaemolyticus

Rep.

Levels of spiking (CFU/g)

Amount of cellsb(CFU/rxn)

LAMP

PCR

   

Fluorescence-based

Turbidity-based

F3/B3

toxR

   

Ct (min)

Mt (°C)

Tt (min)

  

1

5.6 × 108

1.0 × 106

20.61 ± 2.04

82.16 ± 0.05

31.2 ± 2.97

+

+

 

5.6 × 107

1.0 × 105

22.02 ± 2.04

81.36 ± 1.20

35.3 ± 1.13

+

+

 

5.6 × 106

1.0 × 104

25.26 ± 0.56

81.87 ± 0.10

42.55 ± 2.2

+

+

 

5.6 × 105

1.0 × 103

34.58 ± 2.25

82.45 ± 0.23

52.45 ± 2.75

+

-

 

5.6 × 104

1.0 × 102

-

-

-

-

-

 

5.6 × 103

10

-

-

-

-

-

2

1.7 × 108

3.1 × 105

21.78 ± 0.59

82.41 ± 0.11

29.4 ± 0.85

+

+

 

1.7 × 107

3.1 × 104

23.68 ± 0.16

82.25 ± 0.10

33.25 ± 0.35

+

+

 

1.7 × 106

3.1 × 103

29.08 ± 0.45

82.60 ± 0.34

40.4 ± 4.67

+

-

 

1.7 × 105

3.1 × 102

31.77 ± 2.23

82.50 ± 0.18

47.7 ± 1.27

-

-

 

1.7 × 104

31

-

-

-

-

-

 

1.7 × 103

3.1

-

-

-

-

-

3

1.1 × 109

2.0 × 106

20.74 ± 0.03

82.48 ± 0.01

31.25 ± 4.02

+

+

 

1.1 × 108

2.0 × 105

24.14 ± 0.24

82.37 ± 0.05

35.55 ± 3.73

+

+

 

1.1 × 107

2.0 × 104

27.42 ± 0.60

82.48 ± 0.11

40.75 ± 3.88

+

+

 

1.1 × 106

2.0 × 103

33.26 ± 2.84

82.50 ± 0.26

44.8 ± 0.7

+

-

 

1.1 × 105

2.0 × 102

35.57 ± 1.73

82.65 ± 0.09

47.25 ± 0.35

-

-

 

1.1 × 104

20

-

-

-

-

-

  1. Bolded are detection limits by each assay.
  2. aFor each independently prepared template, two times of LAMP reactions were performed and the data presented are means ± standard deviations for the 2 LAMP repeats.
  3. bCFU/reaction was calculated by using CFU/g × 0.09 g/ml × 10 × 2 × 10-3, i.e., CFU/g × 1.8 × 10-3.