LAMP (loop-mediated isothermal amplification) multi-dimensional visual detection color indicator and RNA (ribonucleic acid)/DNA (deoxyribonucleic acid) detection premix solution

A color indicator, multi-dimensional technology, applied in the field of molecular biology, can solve the problems of reducing the speed and accuracy of on-site diagnosis, waiting for a long time to open the aerosol, unable to interpret and other problems. , the effect of easy operation

Pending Publication Date: 2022-04-08
YEASEN BIOTECHNOLOGY (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods all require professional testing equipment, long waiting time and the disadvantages of aerosol pollution after opening the cap.
Detection methods using pH indicators or magnesium ion chelating agents as indicators have developed rapidly in recent years. Compared with detection methods such as electrophoresis, DNA intercalation fluorescent groups, turbidity of magnesium pyrophosphate, and conversion of pyrophosphate, it has rapid judgment, Low pollution rate and other advantages have great advantages in on-site diagnosis, but due to the extremely subtle difference between weak positive and negative results, it is impossible to quickly interpret the results, which reduces the speed and accuracy of on-site diagnosis

Method used

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  • LAMP (loop-mediated isothermal amplification) multi-dimensional visual detection color indicator and RNA (ribonucleic acid)/DNA (deoxyribonucleic acid) detection premix solution
  • LAMP (loop-mediated isothermal amplification) multi-dimensional visual detection color indicator and RNA (ribonucleic acid)/DNA (deoxyribonucleic acid) detection premix solution
  • LAMP (loop-mediated isothermal amplification) multi-dimensional visual detection color indicator and RNA (ribonucleic acid)/DNA (deoxyribonucleic acid) detection premix solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Selection of COVID-19 characteristic nucleic acid sequence and design and synthesis of corresponding LAMP primers

[0086] According to the Gene E Primer Set (NEB, SARS-CoC-2 LAMP Primer Sequence) sequence of COVID-19, 6 corresponding LAMP primers (N2-F3, N2-B3, N2-FIP, N2-BIP, N2-LF, N2-LB); all primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. Primer sequences are shown in Table 1.

[0087]

[0088]

Embodiment 2

[0089] Example 2 Selection of Mycoplasma Characteristic Nucleic Acid Sequence and Design and Synthesis of Corresponding LAMP Primers

[0090] According to the characteristic nucleic acid sequence of Mycoplasma, 6 corresponding LAMP primers (M-F3, M-B3, M-FIP, M-BIP, M-LF, M-LB) were used; all primers were provided by Sangon Bioengineering (Shanghai ) Co., Ltd. on behalf of the synthesis. Primer sequences are shown in Table 2.

[0091]

Embodiment 3

[0092] Example 3 Selection of HBV characteristic nucleic acid sequences and design and synthesis of corresponding LAMP primers

[0093] According to the characteristic nucleic acid sequence of HBV, 6 corresponding LAMP primers (HBV-F3, HBV-B3, HBV-FIP, HBV-BIP, HBV-LF, HBV-LB) were used; all primers were provided by Sangon Bioengineering (Shanghai ) Co., Ltd. on behalf of the synthesis. Primer sequences are shown in Table 3.

[0094]

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Abstract

The invention discloses an LAMP (loop-mediated isothermal amplification) multi-dimensional visual detection color indicator. The LAMP multi-dimensional visual detection color indicator is characterized by comprising phenol red, chrome black T and Eva Green. The invention also discloses an LAMP multi-dimensional visual RNA/DNA (Ribonucleic Acid/Deoxyribose Nucleic Acid) detection premix solution. According to the invention, phenol red is used as a pH indicator, chrome black T is used as a metal ion indicator, Eva Green is used as a fluorescent indicator, and phenol red, chrome black T and Eva Green cooperate with each other in a certain concentration range. According to the method, the low copy number can be detected, the false positive rate is reduced, and the problem that weak positives caused by primer design or too low template concentration are difficult to distinguish during photographing retention or visual distinguishing can be perfectly solved. The color indicator disclosed by the invention has the characteristics of high resolution, high sensitivity, low false positive rate and multi-dimensional verification, and has the effect of judging whether a sample is positive or negative only through naked eyes or 365nm excitation.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a LAMP multi-dimensional visual detection color indicator and an RNA / DNA detection premix. Background technique [0002] Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique. It can be detected by electrophoresis, DNA intercalation of fluorescent groups, turbidity of magnesium pyrophosphate, and conversion of pyrophosphate. However, these methods all require professional testing equipment, long waiting time, and have the disadvantages of aerosol pollution after opening the cap. Detection methods using pH indicators or magnesium ion chelating agents as indicators have developed rapidly in recent years. Compared with detection methods such as electrophoresis, DNA intercalation fluorescent groups, turbidity of magnesium pyrophosphate, and conversion of pyrophosphate, it has rapid judgment, The advantages such as low pollutio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844C12Q1/70C12Q1/689C12N15/11C12R1/93C12R1/35
Inventor 宋东亮罗元廷黄成刘娜曹振
Owner YEASEN BIOTECHNOLOGY (SHANGHAI) CO LTD
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