Streptococcus faecalis powdering LAMP rapid detection kit, and use method thereof

A detection kit, a technology for Streptococcus faecalis, applied in biochemical equipment and methods, microorganism-based methods, and microbial determination/inspection, etc., can solve the problem of indistinguishable results, no mention of sample pretreatment methods, and protective effects. limited and other problems, to achieve the effect of obvious difference in color change, reduce solution configuration steps, and avoid aerosol pollution

Active Publication Date: 2017-03-29
GUANGDONG HUANKAI BIOLOGICAL SCI & TECH CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In practical applications, the corresponding solution reaction system must be prepared before detection, and there are many components in it, which can easily lead to reagent contamination, detection interval contamination, and result errors caused by frequent operations. In addition, some reagents in the reaction system need to be frozen. , For example, the required Bst DNA polymerase needs to be stored strictly at -20°C. If the temperature is too high, it will affect its enzyme activity and cause the detection reagent to fail.
Therefore, the high-temperature long-distance transportation and on-site application of the reagent are inconvenient, and the corresponding cost is also increased.
At the same time, the current detection method has not been popularized in the fields of food safety and medicine, especially in the rapid detection of water bodies. As a non-standard method, the existing LAMP products are only fo

Method used

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  • Streptococcus faecalis powdering LAMP rapid detection kit, and use method thereof
  • Streptococcus faecalis powdering LAMP rapid detection kit, and use method thereof
  • Streptococcus faecalis powdering LAMP rapid detection kit, and use method thereof

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Embodiment Construction

[0035] A rapid detection kit for Streptococcus faecalis in water, comprising a freeze-dried isothermal amplification agent, a complex solution, a positive control dry powder, a negative control, a chromogenic solution, a lysate and a blocking solution, and the freeze-dried isothermal amplification agent contains dNTPs, Bst DNA polymerase, outer primer, inner primer, loop primer and lyoprotectant, the lyoprotectant consists of trehalose, glycine and bovine serum albumin.

[0036] The composition of the lyophilized isothermal amplification agent before lyophilization is: 0.9~1.6mmol / L dNTPs, 0.3~0.4U / μL BstDNA polymerase, 0.1~0.5μmol / L outer primer F3 / B3, 1.0~2.0μmol / L Inner primer FIP / BIP, 0.5-1.5 μmol / L loop primer LF / LB and lyophilized compound protectant containing 3%-6% (w / v) trehalose, 0.01%-0.1% (w / v) glycine, 0.1 %-1% bovine serum albumin, the balance is sterile ultrapure water.

[0037] The composition of the lyophilized isothermal amplification agent before lyophiliza...

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Abstract

The invention discloses a Streptococcus faecalis powdering LAMP rapid detection kit, and a rapid detection method based on the kit. The kit comprises a freeze-dried isothermal amplification agent, a re-dissolving solution, a positive contrast dry powder, a negative contrast substance, a colorimetric solution, a lysate and a sealing solution, and the freeze-dried isothermal amplification agent contains dNTPs, Bst DNA polymerase, an amplimer and a freeze-drying protection agent. The primer in the kit corresponds to eight segments of a Streptococcus faecalis specific gene sequence, the colorimetric solution is added after a reaction, and a negative and positive colorimetric difference is obvious. The kit has the advantages of strong specificity, high sensitivity, easiness in operation and simplicity in result judgment, and is suitable for the quality control and the safe and rapid detection of foods, beverages, water and other raw and auxiliary materials. The kit is convenient for normal temperature transportation, is simple and fast to use without special devices, reduces the cost, widens the application range of products, is suitable for onsite rapid detection and base remote region related detection, and has wide application prospect.

Description

technical field [0001] The invention relates to a dry powdered LAMP rapid detection reagent and a rapid detection method based thereon, in particular to a dry powdered LAMP rapid detection kit for Streptococcus faecalis and a rapid detection method based thereon. Background technique [0002] Streptococcus faecalis is generally distributed in the intestinal tract of humans and animals and is excreted with feces. It is a thermostable bacterium that can still grow well at a temperature of 44.5±0.2°C. As drinking water, the water source is easily polluted directly or indirectly by feces. If there are intestinal infectious diseases in these pollution sources, the water source may contain live pathogenic factors. Contaminated drinking water will cause diseases in the crowd. As an indicator bacterium of sanitary bacteria in drinking water, it is very important in the detection of drinking water sanitation. Streptococcus faecalis can reflect whether the water body is polluted by fe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12R1/46
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2521/101C12Q2527/125
Inventor 万强周杨吴清平卢勉飞曲晓莹杨宁蔡芷荷
Owner GUANGDONG HUANKAI BIOLOGICAL SCI & TECH CO LTD
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