Lactobacillus acidophilus with capability of copolymerizing helicobacter pylori and application of lactobacillus acidophilus
A technology of Lactobacillus acidophilus and microbial strains, applied in the field of microorganisms, can solve the problems of high cost, complicated production principle, and many components, achieve high copolymerization rate, and treat Helicobacter pylori infection
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Embodiment 1
[0091] Embodiment 1: Screening and identification of Lactobacillus acidophilus
[0092] 1. Screening
[0093] Take fresh feces from healthy people as the sample. After pretreatment, the sample is stored in about 30% glycerol in a -80°C refrigerator. After taking out and thawing, mix the sample and add 0.5mL sample to 4.5mL 0.9% normal saline. Gradient dilution, select the appropriate gradient dilution to spread on the MRS solid medium, culture at 37°C for 48 hours, pick typical colonies to streak on the MRS plate for purification, pick a single colony and transfer to liquid MRS liquid medium for enrichment , preserved in 30% glycerol to obtain strain CCFM1208.
[0094] 2. Identification
[0095] The genome of CCFM1208 was extracted, and the 16S rDNA of CCFM1208 was amplified and sequenced (completed by Shanghai Meiji Biomedical Technology Co., Ltd.). After sequencing analysis, the 16S rDNA sequence of the bacterial strain was shown in SEQ ID NO.1. The sequence Comparing in ...
Embodiment 2
[0098] Example 2: Screening of Lactobacillus with co-aggregation with Helicobacter pylori
[0099] Specific steps are as follows:
[0100] (1) Culture of Helicobacter pylori:
[0101] Dissolve Helicobacter pylori on Columbia blood plate (Columbia blood plate: 39g Columbia medium solid powder in 1L water, sterilize at 121°C for 15min, add 7.5% (v / v) sterile solution after cooling to 55°C-60°C Defiberized sheep blood, mixed evenly and then poured onto the plate) was streaked, and placed in a three-gas incubator (85% N) at 37°C 2 , 10% CO 2 , 5% O 2 ) was cultivated for 3 days to obtain a single colony;
[0102] Pick a single colony and inoculate it in BHI liquid medium containing 5% (v / v) fetal bovine serum, in a three-gas incubator at 37°C (85% N 2 , 10% CO 2 , 5% O 2 ) was cultivated for 4 days to obtain the seed liquid; the seed liquid was inoculated in the BHI liquid medium with an inoculum size of 2% (v / v), and in a three-gas incubator at 37°C (85%N 2 , 10% CO 2 , ...
Embodiment 3
[0110] Example 3: Effects of incubation time with Helicobacter pylori on the copolymerization of Lactobacillus acidophilus and Helicobacter pylori
[0111] Specific steps are as follows:
[0112] (1) Culture of Helicobacter pylori:
[0113] Dissolve Helicobacter pylori on Columbia blood plate (Columbia blood plate: 39g Columbia medium solid powder in 1L water, sterilize at 121°C for 15min, add 7.5% (v / v) sterile solution after cooling to 55°C-60°C Defiberized sheep blood, mixed evenly and then poured onto the plate) was streaked, and placed in a three-gas incubator (85% N) at 37°C 2 , 10% CO 2 , 5% O 2 ) was cultivated for 3 days to obtain a single colony; pick a single colony and inoculate it in the BHI medium containing 5% (v / v) fetal bovine serum, in a three-gas incubator at 37°C (85% N 2 , 10% CO 2 , 5% O 2 ) was cultivated for 4 days to obtain seed liquid;
[0114] The seed solution was inoculated in BHI medium with an inoculum size of 2% (v / v), and was incubated i...
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