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Lactobacillus delbrueckii polymerase chain reaction (PCR) detection and identification specific primer pair

A Lactobacillus delbrueckii, specific technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of poor accuracy, long inspection cycle, cumbersome operation, etc.

Inactive Publication Date: 2011-08-10
INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, microbial fertilizer products of Lactobacillus delbrueckii as an effective bacterium are increasing; at the same time, in the process of product quality inspection, it is found that there are many types of Lactobacillus with similar phenotypes, and it is difficult to quickly and accurately distinguish them. Traditional detection methods are mainly based on morphology. The characteristics and various physiological and biochemical methods are mainly used, and there are disadvantages such as long inspection cycle, cumbersome operation, high cost, and poor accuracy.

Method used

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  • Lactobacillus delbrueckii polymerase chain reaction (PCR) detection and identification specific primer pair
  • Lactobacillus delbrueckii polymerase chain reaction (PCR) detection and identification specific primer pair
  • Lactobacillus delbrueckii polymerase chain reaction (PCR) detection and identification specific primer pair

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Experimental program
Comparison scheme
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Embodiment

[0058] Lactobacillus delbrueckii specific primers were used for PCR detection of standard strains.

[0059] (1) Material

[0060] Test bacteria: the standard strains listed in Table 1 below are test bacteria.

[0061] Table 1

[0062]

[0063]

[0064] (2) Design and synthesis of primers

[0065] Lactobacillus delbrueckii specific primers are designed according to the gyrB nucleic acid sequence of Lactobacillus delbrueckii. The primers are shown in Table 2. The primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0066] Table 2

[0067] type

[0068] Lactobacillus delbrueckii

[0069] (3) DNA extraction: Take 1.5 mL of bacterial liquid into an Eppendorf tube, and centrifuge at 13000 r / min for 2 minutes to collect bacterial cells. Wash the collected cells twice with 1×TE buffer solution, then add them to the above-mentioned 2mL centrifuge tube, add about 0.7-1.0g of beads (beater), add 700 μL of phosphate buff...

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Abstract

The invention discloses a pair of specific primers for Lactobacillus delbrueckii, which can quickly and accurately detect Lactobacillus delbrueckii. Meanwhile, the invention establishes a PCR identification method for the Lactobacillus delbrueckii, which can detect and identify Lactobacillus delbrueckii in microorganism manure according to whether an amplification product generates a specific Lactobacillus delbrueckii band (of a size of 182bp).

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, a primer pair for rapid detection and identification of Lactobacillus delbrueckii by PCR technology is provided, which is suitable for application by microbial fertilizer quality inspection institutions and strain identification institutions. Background technique [0002] Lactobacillus delbrueckii (Lactobacillus delbrueckii) is a species of Lactobacillus, including 4 subspecies, of which L.elbrueckii subsp.bulgaricus and L.elbrueckii subs p.delbrueckii are microorganisms Fertilizer commonly used production strains. Like other lactobacilli commonly used in microbial fertilizers, it has good effects in decomposing organic materials, purifying water, promoting growth and inhibiting pathogenic bacteria. [0003] At present, microbial fertilizer products of Lactobacillus delbrueckii as an effective bacterium are increasing; at the same time, in the process of product quality inspec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 杨小红关大伟曹凤明李俊沈德龙姜昕冯瑞华李力陈慧君葛一凡邴晓会
Owner INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI