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A method for rapid detection of Pseudomonas aeruginosa in textiles

A technology of Pseudomonas aeruginosa and textiles, applied in the detection field of Pseudomonas aeruginosa, can solve the problems of cumbersome procedures, inability to detect multiple components at the same time, long cycle, etc.

Active Publication Date: 2020-06-23
INSPECTION & QUARANTINE TECH CENT HENAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional detection methods have cumbersome procedures, long cycles and low sensitivity
Immunological methods are difficult to prepare antibodies, cannot detect multiple components at the same time, require high skills for experimenters, and are prone to cross-contamination
Molecular biology methods have the advantages of high sensitivity, accuracy, and speed, but require special instruments and equipment, are easily polluted, and require high inspection personnel

Method used

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  • A method for rapid detection of Pseudomonas aeruginosa in textiles
  • A method for rapid detection of Pseudomonas aeruginosa in textiles
  • A method for rapid detection of Pseudomonas aeruginosa in textiles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] A method for rapidly detecting Pseudomonas aeruginosa in textiles, comprising the following steps:

[0100] Step 1: Sample enrichment

[0101] Aseptically open the textile sample for inspection, cut the sample evenly with sterile scissors, accurately weigh 25g of the cut sample on an electronic balance, cut it into pieces and add it to a sterile homogeneous bag containing 225mL of SCDLP liquid medium. Beat with a slap-type homogenizer for 1min to 2min, and mix well. Put the prepared samples into a constant temperature incubator at 36°C±1°C, and enrich the bacteria for 24h±2h.

[0102] Step 2: LAMP test

[0103] 2.1) Preparation of DNA template: Take 1mL of SCDLP enrichment solution and put it into a 1.5mL sterile Eppendorf tube, centrifuge at 10000r / min in a high-speed refrigerated centrifuge for 5min, discard the supernatant; add 1mL of sterilized double-distilled water, mix well, and centrifuge in a high-speed refrigerated centrifuge Centrifuge at 10,000r / min in th...

Embodiment 2

[0136] A method for rapidly detecting Pseudomonas aeruginosa in textiles, comprising the following steps:

[0137] Step 1: Sample enrichment

[0138] Aseptically open the textile sample for inspection, cut the sample evenly with sterile scissors, accurately weigh 25g of the cut sample on an electronic balance, cut it into pieces and add it to a sterile homogeneous bag containing 225mL of SCDLP liquid medium. Beat with a slap-type homogenizer for 1min to 2min, and mix well. Put the prepared samples into a constant temperature incubator at 36°C±1°C, and enrich the bacteria for 24h±2h.

[0139] Step 2: LAMP test

[0140] 2.1) Preparation of DNA template: Take 1mL SCDLP enrichment solution and put it into a 1.5mL sterile Eppendorf tube, centrifuge in a high-speed refrigerated centrifuge at 10,000r / min for 5min, discard the supernatant; add 1mL sterilized double-distilled water to mix, and freeze at high speed Centrifuge at 10,000r / min in a centrifuge for 5min, discard the super...

Embodiment 3

[0172] A method for rapidly detecting Pseudomonas aeruginosa in textiles, comprising the following steps:

[0173] Step 1: Sample enrichment

[0174] Aseptically open the textile sample for inspection, cut the sample evenly with sterile scissors, accurately weigh 25g of the cut sample on an electronic balance, cut it into pieces and add it to a sterile homogeneous bag containing 225mL of SCDLP liquid medium. Beat with a slap-type homogenizer for 1min to 2min, and mix well. Put the prepared samples into a constant temperature incubator at 36°C±1°C, and enrich the bacteria for 24h±2h.

[0175] Step 2: LAMP test

[0176] 2.1) Preparation of DNA template: Take 1mL of SCDLP enrichment solution and put it into a 1.5mL sterile Eppendorf tube, centrifuge at 10000r / min in a high-speed refrigerated centrifuge for 5min, discard the supernatant; add 1mL of sterilized double-distilled water, mix well, and centrifuge in a high-speed refrigerated centrifuge Centrifuge at 10,000r / min in th...

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Abstract

The invention discloses a method for rapidly detecting pseudomonas aeruginosa in a textile. The method comprises designing a set of LAMP specific primers based on the pseudomonas aeruginosa specific ETA conserved sequence as target gene sequence, carrying out enrichment culture on a textile sample, carrying out product LAMP amplification to obtain white precipitates, comparing and observing the precipitates to determine a result or observing a result through a turbidity meter, wherein when the sample reaction turbidity value is less than 0.1, the sample result is negative and when the sample reaction turbidity value is greater than or equal to 0.1, the sample result is positive, separating the positive sample enrichment fluid inoculation flat plate, picking a typical or suspicious colony on the flat plate, and carrying out fast identification through a MALDI-TOF-MS technology to obtain an identification result. The method has the advantages of simple operation, fastness, high efficiency, strong specificity, high sensitivity and convenient and visual observation.

Description

technical field [0001] The invention relates to a method for detecting Pseudomonas aeruginosa, in particular to a method for rapidly detecting Pseudomonas aeruginosa in textiles. Background technique [0002] Pseudomonas aeruginosa (P.Aeruginosa), formerly known as Pseudomonas aeruginosa, is a highly lethal, hemolytic, and drug-resistant Gram-negative bacillus, which belongs to Pseudomonas in the Pseudomonas family in classification There are more than 200 kinds of bacteria in this genus, which are widely distributed in nature and are one of the most common bacteria in soil. The most important growth condition of this bacterium is a humid environment, and other conditions are not very demanding. Textiles are highly hygroscopic and have all the factors for the growth of Pseudomonas aeruginosa, which can provide an ideal environment for its reproduction, including suitable pH, temperature and water nutrition, and can also provide the large surface area required for its reprod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6844C12Q1/689C12Q1/04G01N33/68C12R1/385
CPCC12Q1/6844C12Q1/689G01N33/6851G01N2333/21C12Q2531/119C12Q2545/113C12Q2563/107
Inventor 李轲郭会清郭华麟张淑霞徐超乔晴禹建鹰张超峰
Owner INSPECTION & QUARANTINE TECH CENT HENAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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