Method for rapidly detecting listeria monocytogens and staphylococcus aureus
A technology for Listeria monocytogenes and Staphylococcus monocytogenes is applied in the field of rapid detection of Listeria monocytogenes and Staphylococcus aureus in food, which can solve the problems of complicated operations and steps, endangering consumer safety, and low detection rate and content. , to reduce the interference of detection accuracy, avoid false negative phenomenon, and achieve the effect of high sensitivity
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Embodiment 1
[0031] The screening of embodiment 1 BASV concentration
[0032] 1.1 Listeria monocytogenes
[0033] Take 225mL of PBS buffer solution containing different concentrations of BASV (0.02mol / L in PBS) in 6 parts respectively in homogenization bags, add 25g samples containing Listeria monocytogenes to each homogenization bag and homogenize for 30s, add The concentrations of BASV were 0% (no BASV added), 0.5%, 1%, 5%, 8% and 10%.
[0034] In addition, 225 mL of Listeria monocytogenes broth LB1 was taken as a comparison solution, and 25 g of a sample containing Listeria monocytogenes was added and homogenized for 30 seconds to prepare a sample solution as a comparison example.
[0035] According to the traditional detection method, operations such as enrichment, streak separation, and identification were performed on the above sample liquid, and the test results were obtained. The results are shown in Table 1.
[0036] Staphylococcus aureus
[0037] With reference to the above-me...
Embodiment 2
[0041] Example 2 Optimum Addition of Growth Solution
[0042]Take 25 g of positive samples containing Listeria monocytogenes and Staphylococcus aureus respectively, add them to a homogenizing bag containing 1% BASV in 225 mL of PBS buffer, and homogenize for 30 seconds. Take 6 parts of 5mL sample bacterial liquid, collect the bacterial cells after centrifugation at 4°C and 8000rpm, add 0.1mL, 0.5mL, 1mL, 2mL, 4mL, 5mL of growth solution, and mix well. Then smear it on the plates corresponding to the two kinds of bacteria respectively, carry out operations such as streak separation and biochemical test according to GB4789, and obtain the test results, as shown in Table 2. The whole process detection method of GB4789 is also used as a comparison in the table.
[0043] Table 2 The results of colony detection with different amounts of growth solution added
[0044]
[0045] It can be seen from the table that with the increase of the growth liquid, the number of bacteria also i...
Embodiment 3
[0046] Example 3 Optimum time for ultrasonic oscillation mixing
[0047] Take 25 g of positive samples containing Listeria monocytogenes and Staphylococcus aureus, add them to homogenizing bags containing 1% BASV in 225 mL of PBS buffer, and homogenize for 30 seconds. Take 5 parts of 5mL sample bacterial liquid, collect the bacterial cells after centrifugation at 4°C and 8000rpm, add 1mL growth liquid, mix the growth liquid with the bacterial liquid, and ultrasonically oscillate respectively. The oscillation time of the five samples is 1min, 3min, 5min, and 8min in turn. and 10min, after shaking, spread on the plates corresponding to the two kinds of bacteria to carry out operations such as streak separation and culture, and the detection results are shown in Table 3.
[0048] Table 3 The number of colonies detected by ultrasonic oscillation at different times
[0049]
[0050] The results showed that when the shaking mixing time was 5 minutes, the number of colonies detec...
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