A method for rapid detection of Salmonella in fresh milk
A technology for salmonella and fresh milk, which is applied in the field of microbial detection, can solve problems such as time-consuming, false negative results, and low detection rate, and achieve the effects of shortening the detection time, increasing the detection rate, and solving detection problems
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Embodiment 1
[0027] Example 1 Screening test
[0028] 1.1 Selection of fresh milk sample buffer
[0029] Take 4 copies of 25mL fresh milk Salmonella positive samples and add them to homogeneous bags containing 225mL buffer respectively. The buffers in the homogeneous bags are: ddH 2 O, 0.01mol / L PBS buffer (pH7.4, the same below), carbonate buffer, buffered peptone water (BPW). Then, according to the traditional salmonella detection method, the above-mentioned fresh milk mixture was enriched, streaked and separated, and biochemical tests were carried out. The test results are shown in Table 1.
[0030]
[0031] It can be seen from Table 1 that both BPW and 0.01mol / L PBS can effectively protect Salmonella in samples, but the cost of BPW is high, so 0.01mol / L PBS was selected as the pretreatment buffer.
[0032] 1.2 Comparison of the detection rate between the double filter method and the national standard method
[0033] Take 25mL of positive fresh milk samples, put them into a steril...
Embodiment 2
[0048] A kind of rapid detection method of Salmonella in fresh milk, comprises the following steps:
[0049] 1) Aseptically take 25 mL of fresh milk sample, add it into a sterile homogeneous bag containing 25 mL of phosphate-buffered saline solution containing 2 g of cellulose resin, and prepare a 50 mL fresh milk mixed solution. Homogenizer beating for 2min;
[0050]2) Rinse the mixed solution of fresh milk with 200mL PBS and quickly filter it into the container with filter paper, centrifuge the filtrate at a centrifugation speed of 6000r / min and a centrifugation temperature of 4°C for 10min, and filter the supernatant with a 0.22μm microporous membrane. Concentrate microorganisms on microporous membranes;
[0051] 3) Cut the microporous membrane into pieces with sterile scissors, rinse and soak for 2 minutes with a mixture of 5ml TTB enrichment solution and 5ml PBS buffer solution to obtain a concentrated bacterial suspension;
[0052] 4) Take 100 μL of Salmonella immune m...
Embodiment 3
[0056] Embodiment 3 comparative test
[0057] Set the concentration to 1 x 10 9 The CFU / mL Salmonella bacterial liquid was diluted according to a certain gradient, and 1 mL of the diluted solution was added to 25 mL of fresh milk samples to prepare fresh milk positive samples. According to the method described in Example 2, and at the same time, the "GB 4789.4-2010" national standard method was used for comparison, and the fresh milk positive sample and the fresh milk sample were detected. The comparison test results are shown in Table 5.
[0058]
[0059] It can be seen from Table 5 that, compared with the national standard method, the detection method of the present invention has higher sensitivity and greatly improves the detection rate of Salmonella in fresh milk samples. In addition, for this group of tests, the national standard method takes 4-7 days to detect a sample, and the method of the present invention takes 50 minutes to detect a sample, and the latter greatl...
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