CN-122016739-A - LAMP rapid fluorescence detector based on multiple fluorescence detection reagent

CN122016739ACN 122016739 ACN122016739 ACN 122016739ACN-122016739-A

Abstract

The invention belongs to the technical field of nucleic acid isothermal amplification detection and discloses an LAMP rapid fluorescence detector based on multiple fluorescence detection reagents, which comprises a shell, wherein a laser light source module, a reaction test tube and an optical beam splitter module are arranged in the shell, the optical beam splitter module is used for splitting an excitation beam into at least two independent sub-beams to form at least two optical path channels, an optical path switch module and an optical filter module are sequentially arranged on each optical path channel, the sub-beams deviating from the original propagation direction change the propagation direction through the reflector module, and the beams are guided to the reaction test tube, and when the detection is carried out, the optical path switch module is used for controlling the on-off of the optical path channels, so that at any detection moment, only one optical path channel is in a beam path state, and the rest optical path channels are in a beam blocking state. The invention simplifies the detection steps while remarkably reducing the manufacturing cost, meets the high-performance requirement of on-site rapid detection, and expands the use scene.

Inventors

ZHAO QIAN
SONG SHUISHAN
LI BAOLU
LI XIN
JIA YINGXIN
JIA ZHENHUA
LU MINGYUE
LI LIU

Assignees

河北省科学院生物研究所
河北省机电一体化中试基地有限公司

Dates

Publication Date: 20260512
Application Date: 20251219

Claims (8)

1. LAMP rapid fluorescence detector based on multiple fluorescence detection reagent, its characterized in that includes the casing, is provided with in the casing: the laser light source module is used for providing an excitation light beam; The reaction test tube is used for accommodating the LAMP reaction system and the fluorescent detection reagent; The optical beam splitter module is used for splitting the excitation beam into at least two paths of independent sub-beams to form at least two optical path channels, and each optical path channel is sequentially provided with: the optical path switch module is used for controlling the on-off of the corresponding optical path channel; the optical filter module is used for adjusting the wavelength of the corresponding sub-beam to a preset value; the sub-beams deviating from the original propagation direction change the propagation direction through the reflector module, guide the beams to the reaction test tube so as to excite fluorescent signals in the reaction test tube, and control the on-off of the light path channels through the light path switch module when detecting, so that only one light path channel is in a light beam path state at any detection moment, and the rest light path channels are in a light beam blocking state.
2. The LAMP rapid fluorescence detector based on the multiple fluorescence detection reagent according to claim 1, wherein the optical beam splitter module comprises a primary optical beam splitter and a secondary optical beam splitter; The first-stage optical beam splitter splits an incident beam from the laser source module into a first transmitted beam and a first reflected beam according to a first preset proportion, and the second-stage optical beam splitter splits the first transmitted beam into a second transmitted beam and a second reflected beam according to a second preset proportion.
3. The LAMP rapid fluorescence detector based on the multiple fluorescence detection reagent according to claim 2, wherein the optical channel formed by the first reflected light beam and the second reflected light beam is converged to the reaction cuvette through the convex lens module.
4. The LAMP rapid fluorescence detector based on multiple fluorescence detection reagent according to claim 2 or 3, wherein the first preset ratio is 70:30 and the second preset ratio is 50:50.
5. The LAMP rapid fluorescence detector based on multiple fluorescence detection reagents according to claim 4, wherein the light path switch module comprises an electrically controlled dimming glass and a control switch.
6. The LAMP rapid fluorescence detector based on multiple fluorescence detection reagents according to claim 5, wherein the filter module is detachably arranged in the housing and is replaced according to the multiple fluorescence detection reagents used.
7. The LAMP rapid fluorescence detector based on multiple fluorescence detection reagents according to any one of claims 1 to 3, 5 and 6, wherein an observation window is formed in the housing at a position corresponding to the reaction tube for observing the change of fluorescence color in the reaction tube.
8. The rapid LAMP fluorescence detector based on multiple fluorescence detection reagents according to claim 7, wherein the laser light source module is a light emitting diode.

Description

LAMP rapid fluorescence detector based on multiple fluorescence detection reagent Technical Field The invention belongs to the technical field of nucleic acid isothermal amplification detection, and relates to an LAMP rapid fluorescence detector, in particular to an LAMP rapid fluorescence detector based on multiple fluorescence detection reagents. Background The loop-mediated isothermal amplification (LAMP) technology is used as a high-efficiency nucleic acid isothermal amplification method, has the remarkable advantages of high reaction speed (usually completed within 15-60 minutes), simple operation (without complex thermal cycling equipment), high specificity (using 4-6 specific primers), visual judgment of byproduct magnesium pyrophosphate precipitation turbidity or fluorescent signals and the like, and has great application potential in the fields of pathogen diagnosis, food safety monitoring, environmental microorganism detection and the like, particularly in the field rapid detection scene. By combining the LAMP technology with fluorescence detection, the real-time and quantitative monitoring of the amplification process can be realized by adding fluorescent dye (such as SYBR Green I) specifically combined with the amplification product or a sequence-specific fluorescent probe (such as a quenched fluorescent probe) into the reaction system, and the sensitivity, accuracy and objectivity of the detection are greatly improved. In particular to a multiplex fluorescent LAMP detection technology, by using probes aiming at different targets and marked with fluorescent groups with different emission wavelengths, a plurality of target nucleic acid sequences can be detected simultaneously in a single reaction tube, the detection flux and the detection efficiency are obviously improved, and the multiplex fluorescent LAMP detection technology has important significance for complex infectious pathogen typing, multiplex detection kit development and the like. However, the realization of efficient and reliable multiplex fluorescent LAMP detection is highly dependent on the matched detection instrument. Currently, prior art real-time fluorescence detection devices typically employ a broad spectrum light source (e.g., high pressure mercury lamp, pulsed xenon lamp) in combination with a filter wheel or monochromator to provide excitation light at different wavelengths. Although the light source can cover a wider excitation spectrum and meet the requirements of various fluorescent dyes, the following inherent defects seriously restrict the application in the field of rapid and portable detection, in particular the application of the adaptive multiplex LAMP reagent: (1) The high-pressure mercury lamp, the xenon lamp and the matched power supply and heat dissipation system thereof have larger volume and weight and power consumption of more than tens of watts, so that the whole machine is difficult to miniaturize and lighten, is seriously dependent on stable commercial power supply, and cannot meet the use requirements of the field, vehicle-mounted and basic-level clinics and other environments without stable power supply or movement; (2) To separate excitation light of a specific wavelength from a broad spectrum light source, a complex filter wheel or grating system is required, which not only increases the complexity and cost of the mechanical structure, but also results in light path energy loss. And the excitation wavelength combination is usually fixed or limited and adjustable, so that flexible and accurate matching is difficult to carry out according to the fine difference of the optimal excitation wavelengths of the fluorophores in different multiple fluorescence detection reagents, and the detection sensitivity is influenced. Disclosure of Invention The invention aims to provide an LAMP rapid fluorescence detector based on multiple fluorescence detection reagents, so as to achieve the purposes of rapid detection, applicability to various application scenes and improvement of detection sensitivity. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: The LAMP rapid fluorescence detector based on the multiple fluorescence detection reagents comprises a shell, wherein the shell is internally provided with: the laser light source module is used for providing an excitation light beam; The reaction test tube is used for accommodating the LAMP reaction system and the fluorescent detection reagent; The optical beam splitter module is used for splitting the excitation beam into at least two paths of independent sub-beams to form at least two optical path channels, and each optical path channel is sequentially provided with: the optical path switch module is used for controlling the on-off of the corresponding optical path channel; the optical filter module is used for adjusting the wavelength of the corresponding sub-beam to a preset value; the sub-beams deviating from the o