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Method for quickly detecting beta-hemolytic streptococcus in textile product

A technology for hemolytic streptococcus and textiles, which is applied in the detection field of beta-hemolytic streptococcus, can solve problems such as complex components, and achieve the effects of high sensitivity, wide application range and low detection cost

Inactive Publication Date: 2018-01-30
郭会清
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition of textile samples is complex, and traditional detection methods are prone to introduce too many errors, resulting in false negative results. Therefore, according to the characteristics of textile samples, IMS and RPA technology are scientifically combined to establish a specific enrichment, rapid, simple and accurate detection method , provides a new development direction for the detection of β-hemolytic streptococci, and is expected to become a simple routine detection method, especially suitable for grass-roots inspection and quarantine institutions, which is of great significance for improving the hygiene level of products, ensuring the personal safety of people and promoting the development of international textile trade. Significance

Method used

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  • Method for quickly detecting beta-hemolytic streptococcus in textile product
  • Method for quickly detecting beta-hemolytic streptococcus in textile product
  • Method for quickly detecting beta-hemolytic streptococcus in textile product

Examples

Experimental program
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Effect test

Embodiment 1

[0080] A method for rapidly detecting β-hemolytic streptococci in textiles, comprising the following steps:

[0081] Step 1. Preparation of immunomagnetic beads

[0082] Take 0.1mL of magnetic bead solution in a centrifuge tube filled with 1.0mL of PBST buffer, mix well, centrifuge at 3000r / min for 2min, discard the liquid; repeat the above steps to wash 3 times, add 1.0mL of PBST buffer to resuspend the magnetic beads ; Add 0.1mL of β-hemolytic streptococcus immune serum to the above resuspension, put it on a shaker, and gently rotate and oscillate at room temperature for 3h; wash with 1.0mL 1×washing solution for 5 times, add 0.5mL of resuspension, and in 4 Store at ℃ for later use;

[0083] Step 2. RPA primer and probe design

[0084] Download the target gene sequence encoding HtrA from the NCBI database, use CLUSTAL X software for sequence alignment, and find a specific sequence in the HtrA gene that is only conserved in groups A, B, C, and G of β-hemolytic streptococcus...

Embodiment 2

[0104] A method for rapidly detecting β-hemolytic streptococci in textiles, comprising the following steps:

[0105] Step 1. Preparation of immunomagnetic beads

[0106] Take 0.1mL of magnetic bead solution in a centrifuge tube filled with 1.0mL of PBST buffer, mix well, centrifuge at 3000r / min for 2min, discard the liquid; repeat the above steps to wash 3 times, add 1.0mL of PBST buffer to resuspend the magnetic beads ; Add 0.1mL of β-hemolytic streptococcus immune serum to the above resuspension, put it on a shaker, and gently rotate and oscillate at room temperature for 3h; wash with 1.0mL 1×washing solution for 5 times, add 0.5mL of resuspension, and in 4 Store at ℃ for later use;

[0107] Step 2. RPA primer and probe design

[0108] Download the target gene sequence encoding HtrA from the NCBI database, use CLUSTAL X software for sequence alignment, and find a specific sequence in the HtrA gene that is only conserved in groups A, B, C, and G of β-hemolytic streptococcus...

Embodiment 3

[0128] A method for rapidly detecting β-hemolytic streptococci in textiles, comprising the following steps:

[0129] Step 1. Preparation of immunomagnetic beads

[0130] Take 0.1mL of magnetic bead solution in a centrifuge tube filled with 1.0mL of PBST buffer, mix well, centrifuge at 3000r / min for 2min, discard the liquid; repeat the above steps to wash 3 times, add 1.0mL of PBST buffer to resuspend the magnetic beads ; Add 0.1mL of β-hemolytic streptococcus immune serum to the above resuspension, put it on a shaker, and gently rotate and oscillate at room temperature for 3h; wash with 1.0mL 1×washing solution for 5 times, add 0.5mL of resuspension, and in 4 Store at ℃ for later use;

[0131] Step 2. RPA primer and probe design

[0132] Download the target gene sequence encoding HtrA from the NCBI database, use CLUSTAL X software for sequence alignment, and find a specific sequence in the HtrA gene that is only conserved in groups A, B, C, and G of β-hemolytic streptococcus...

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Abstract

The invention discloses a method for quickly detecting beta-hemolytic streptococcus in a textile product. The method is characterized in that two technologies of IMS (immunomagnetic separation) and RPA (recombinase ploymerase amplification) are combined, and the beta-hemolytic streptococcus in a sample is enriched by an immunomagnetic bead capturing technology; one set of specific primer and probeusing the RPA technology as the core technology is designed for the target gene with the specificity HtrA of the beta-hemolytic streptococcus, and the enriched beta-hemolytic streptococcus is detected by the RPA technology. The method has the advantages that the culture is not needed in the whole process; the time is shortened, the operation is quick, the efficiency is high, the specificity is good, the sensitivity is high, the anti-interference property is strong, the application range is broad, and the detection cost is low; the method is not only suitable for sanitation health and epidemicprevention departments and health and supervision departments, and but also suitable for the sanitation detection in enterprises and families.

Description

technical field [0001] The invention relates to a method for detecting beta-hemolytic streptococcus, in particular to a method for rapidly detecting beta-hemolytic streptococcus in textiles. Background technique [0002] β-hemolytic streptococcus (β-hemolytic streptococcus) is widely distributed in nature and exists in water, air, dust, feces, and the oral cavity, nasal cavity, and throat of healthy humans and animals. Streptococci related to environmental hygiene and personal safety in the genus Streptococcus are mainly β-hemolytic streptococci, which can be divided into 18 groups according to their C antigens, most of which are pathogenic to humans belong to Group A, Group B, and Group C Group and Group G. β-hemolytic streptococcus is highly virulent and invasive to humans, and can produce a variety of extracellular enzymes and exotoxins, causing various suppurative diseases, pneumonia, wound infection, and sepsis. It can be transmitted through direct contact, air drople...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844C12Q1/14
Inventor 李轲郭会清禹建鹰张淑霞郭华麟连素梅徐超苗丽
Owner 郭会清
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