Bacteroides fragilis and applications thereof

A technology of Bacteroides fragilis and strains, which is applied in the fields of food and daily chemical products, medicines, microorganisms, and health care products, can solve the problems of weak clinical curative effect, and achieve the effect of remarkable curative effect and strong stomach acid ability

Active Publication Date: 2017-02-15
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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  • Bacteroides fragilis and applications thereof
  • Bacteroides fragilis and applications thereof
  • Bacteroides fragilis and applications thereof

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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PUM

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Abstract

The invention relates to Bacteroides fragilis and applications thereof, particularly to Bacteroides fragilis ZY-312 having the preservation number of CGMCC No.10685, and applications of the Bacteroides fragilis ZY-312 in preparation of medicines, pharmaceutical compositions, foods, health products and food additives for prevention and / or treatment of antibiotic-associated diarrhea, wherein the Bacteroides fragilis ZY-312 is preserved in the China general microbiological culture collection center on April 2, 2015, has the preservation number of CGMCC No.10685, does not contain enterotoxin gene bft, has characteristics of significantly-improved cholate resistance and significantly-improved gastric acid resistance compared to the existing Bacteroides fragilis, and can effectively prevent and / or treat antibiotic-associated 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, referred to as PMC) is a special type of serious condition with pseudomembrane formation in the colonic mucosa. If not given timely and reasonable treatment, it may cause c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P1/12A23L33/135C12R1/01
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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