Method for identifying bifidobactirium

A technology of bifidobacteria and analysis method, which is applied in the field of molecular biology to identify bifidobacteria, and can solve the problems of distinguishing species, difficult bacteria, etc.

Active Publication Date: 2011-06-29
SHANGHAI JIAODA ONLLY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the 16S rRNA gene sequence is highly conserved, there is little difference in the 16S rRNA gene sequence between certain species of Bifidobacteria, so it is difficult to clearly distinguish bacteria into sp

Method used

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  • Method for identifying bifidobactirium
  • Method for identifying bifidobactirium
  • Method for identifying bifidobactirium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The identification of embodiment 1 Bifidobacterium lactis BL99

[0061] After the Bifidobacterium lactis strain BL99 was anaerobically cultured in the MRS liquid medium for 24 hours, 1ml of the bacterial liquid was taken to extract genomic DNA, and the DNA of the bacterial strain was used as a template, and the genus-specific primer pair Bif500F & Bif500R was used as primers for PCR amplification. Strain BL99 can be amplified to obtain a target fragment of about 500bp, indicating that the strain belongs to the genus Bifidobacterium; at the same time, primer pairs Bif900F & Bif900R and Pk300F & Pk00R were used for PCR amplification, and the electrophoresis of the amplified product is as follows figure 1 shown.

[0062] The obtained three target gene fragments were purified and sequenced respectively, and the following results were obtained:

[0063] Alignment results of target fragment 1 (Bif500 primer pair): the strains with the highest homology include Bifidobacterium...

Embodiment 2

[0067] Identification of Example 2 Bifidobacterium longum BL88-Onlly

[0068] After the Bifidobacterium longum strain BL88 was anaerobically cultured in the MRS liquid medium for 24 hours, 1ml of the bacterial liquid was taken to extract genomic DNA, and the DNA of the bacteria was used as a template, and the genus-specific primer pair Bif500F & Bif500R was used as primers for PCR amplification. The strain BL88-Onlly can be amplified to obtain a target fragment of about 500bp, indicating that the strain belongs to the genus Bifidobacterium. Carry out PCR amplification with primer pair Bif900F & Bif900R and Pk300F & Pk00R simultaneously, the electrophoresis figure of amplified product is as follows figure 1 shown.

[0069] The obtained three target gene fragments were purified and sequenced respectively, and the following results were obtained:

[0070] Alignment results of target fragment 1 (Bif500 primer pair): the strains with the highest homology include Bifidobacterium l...

Embodiment 3

[0074] Example 3 Identification of Bifidobacterium breve BB8

[0075] After the Bifidobacterium breve strain BB8 was anaerobically cultured in the MRS liquid medium for 24 hours, 1 ml of the bacterial liquid was taken to extract genomic DNA, and the DNA of the bacteria was used as a template, and the genus-specific primer pair Bif500F & Bif500R was used as primers for PCR amplification. Strain BB8 can be amplified to obtain a target fragment of about 500bp, indicating that the strain belongs to the genus Bifidobacterium. Simultaneously carry out PCR amplification with primer pair Bif900F & Bif900R and Pk300F & Pk00R, the electrophoresis figure of amplified product is as follows figure 1 shown.

[0076] The obtained three target gene fragments were purified and sequenced respectively, and the following results were obtained:

[0077] Alignment results of target fragment 1 (Bif500 primer pair): the most homologous strain of Bifidobacterium breve B.breve. The target fragment s...

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Abstract

The invention relates to a method for identifying bifidobactirium, in particular to a novel molecular biology method for identifying bifidobactirium. According to the method, PCR (Polymerase Chain Reaction) amplification and sequencing analysis are performed on two target gene segments on bifidobactirium 16SrDNA and one target gene segment on pyruvate kinase coded gene, so that the bifidobactirium strain can be identified on a seed level. The invention provides a more accurate practical method which is much easier for operation for identifying bifidobactirium and is favorable for sorting and research works of the bifidobactirium strains.

Description

technical field [0001] The invention relates to bifidobacteria, in particular to a molecular biology method for identifying bifidobacteria. More precisely, the present invention provides a PCR (Polymerase Chain Reaction, polymerase chain reaction, referred to as PCR) amplification method for two target segments on the bifidobacterium 16S rRNA gene and a target segment on the pyruvate kinase gene. Augmentation and sequencing analysis, a method for the identification of bifidobacterial strains to the species level. Background technique [0002] Bifidobacterium is a Gram-positive bacterium, an obligate anaerobic bacterium with polymorphic characteristics, but overall it has a typical morphology of "branched". According to the description in "Bergey's Bacterial Identification Manual", the genus Bifidobacterium belongs to the class Actinobacteria, the order Bifidobacteriales, and the family Bifidobacteriaceae. In 1899, Dr. Tisser of the Institut Pasteur in France discovered Bif...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张旻杭晓敏张和春陈培青
Owner SHANGHAI JIAODA ONLLY CO LTD
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