A kind of Bacteroides fragilis and its application

A technology for Bacteroides fragilis and Clostridium difficile infection, which is applied in the fields of microorganisms, health care products, food and daily chemical products, and pharmaceuticals. It can solve the problems of weak clinical curative effect and achieve significant curative effect and strong stomach acid ability.

Active Publication Date: 2019-07-05
GUANGZHOU ZHIYI PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After nearly half a century of practice, Europe, North America, Japan, and China are all using probiotic regulators as yogurt fermented beverages, health foods, and medicines. This is a generally recognized fact. However, these preparations mainly contain bifidobacteria and Lactobacillus, after long-term and large-scale application, it is found that its clinical curative effect is getting weaker and weaker, so it is urgent to intensify research and development efforts to further enrich the varieties of probiotics

Method used

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  • A kind of Bacteroides fragilis and its application
  • A kind of Bacteroides fragilis and its application
  • A kind of Bacteroides fragilis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Isolation and purification of Bacteroides fragilis ZY-312

[0042] Reagents and instruments:

[0043] (1) Medium A: Bacteroides-bile-escin (BBE) agar (Qingdao Haibo Biotechnology Co., Ltd., product number: HB7028) was added with an improved formula, and the specific components are shown in Table 1:

[0044] Table I

[0045]

[0046] (2) Medium B: Bacteroides-bile-escin (BBE) agar (Qingdao Haibo Biotechnology Co., Ltd., product number: HB7028) was added with an improved formula, and the specific ingredients are as follows in Table 2:

[0047] Table II

[0048]

[0049] (3) Medium C: Fetal bovine serum was added to Brinell Broth (Qingdao Haibo Biotechnology Co., Ltd., product number: HB0241), and the addition amount was 5% (v / v) (Zhejiang Tianhang Biotechnology Co., Ltd., Brand: Sijiqing, Item No.: HB0205).

[0050] (4) Experimental equipment

[0051] 2.5L sealed culture tank (Mitsubishi Gas Chemical Co., Ltd., C-31)

[0052] Constant temperature incubator (Sh...

Embodiment 2

[0090] Identification of Bacteroides fragilis ZY-312

[0091] The polymerase chain reaction (PCR) primer (synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.) sequence is as follows:

[0092] Primer pair 1:

[0093] Forward primer: 5'-ACGCTTGCACCCCTCCGTATTA-3'

[0094] Reverse primer: 5'-GCTTAGAGTTTGATCCTGGCTCAG-3'

[0095] Primer pair 2:

[0096] Forward primer: 5'-TGGGTGGTTGCTGCCTGGACACA-3'

[0097] Reverse primer: 5'-CATCCGGGTATGGATATGAA-3'

[0098] Primer pair 3:

[0099] Forward primer: 5'-GATGCTCCAGTTACAGCTTCCATTG-3'

[0100] Reverse primer: 5'-CGCCCAGTATATGACCTAGTTCGTG-3'

[0101] bft gene primer pair:

[0102] Forward primer: 5'-GACGGTGTATGTGATTTGTCTGAGAGA-3'

[0103] Reverse primer: 5'-ATCCCTAAGATTTATTATCCCAAGTA-3'

[0104] 1. PCR identification (PCR is the polymerase chain reaction, which is a commonly used method for rapidly amplifying genes)

[0105] (1) 16S rRNA sequence determination

[0106] The above strains were inoculated on medium A, and cult...

Embodiment 3

[0128] Tolerance of Bacteroides fragilis ZY-312 to Gastric Acid

[0129] (1) Artificial gastric juice preparation (according to the 2010 "Chinese Pharmacopoeia" artificial gastric juice preparation method)

[0130] Dissolve 23.4mL of concentrated HCl in 100mL of purified water to obtain dilute hydrochloric acid. Take 8.2mL dilute hydrochloric acid, add 400mL purified water and 5g pepsin ( Porcine source, 1:15000), dilute to 500mL. Stir overnight at 37°C with magnetic force to obtain artificial gastric juice.

[0131] (2) Cell preparation

[0132] Collect the bacterial solution, centrifuge, discard the supernatant, resuspend in normal saline, centrifuge again, discard the supernatant, and keep the bacteria for later use.

[0133] (3) adding artificial gastric juice to determine the number of viable bacteria

[0134] Add artificial gastric juice to the bacteria, resuspend, and measure the number of viable bacteria at 0, 1, 2, and 3 hours respectively.

[0135] The 10-fold...

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Abstract

A kind of Bacteroides fragilis and its application, specifically related to Bacteroides fragilis ZY-312 with the preservation number of CGMCC No.10685, and the preparation of Bacteroides fragilis ZY‑312 for preventing and / or treating antibiotic-associated diarrhea, and a pharmaceutical composition , food, health products and food additives, wherein, the Bacteroides fragilis ZY‑312 was deposited in the China General Microorganism Culture Collection Management Center on April 2, 2015, and its preservation number is CGMCC No.10685. The Bacteroides fragilis ZY‑312 of the present invention does not contain the enterotoxin gene bft, has significantly higher tolerance to bile salts and gastric acid than the existing Bacteroides fragilis, and can effectively prevent and / or treat antibiotic-related diarrhea.

Description

technical field [0001] The invention relates to the technical fields of microorganisms, medicines, health products, food and daily chemical products, in particular to a new strain of Bacteroides fragilis and its application in preventing and / or treating antibiotic-associated diarrhea. Background technique [0002] With the widespread use of antibiotics, antibiotic-associated diarrhea (AAD) has attracted increasing attention. Antibiotic-associated diarrhea refers to diarrhea secondary to the use of antibiotics. It is a relatively common adverse drug reaction, and its incidence varies with different antibiotics, about 5% to 39%. According to the severity of AAD, antibiotic-associated diarrhea includes simple diarrhea, colitis, or pseudomembranous colitis. Pseudomembranous colitis (Pseudomembranous colitis, PMC for short) refers to a special type of serious condition with pseudomembrane formation in the colonic mucosa. If not given timely and reasonable treatment, it may cause...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/741A61P1/12A23L33/135C12R1/01
CPCA61K35/74C12N1/20C12N1/205C12R2001/01
Inventor 智发朝白杨刘洋洋王晔王从峰
Owner GUANGZHOU ZHIYI PHARMA INC
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