Detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphism through single reaction hole

Abstract

The invention discloses a detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphism through a single reaction hole, and belongs to the technical field of detection kits.The detection kit comprises a kit body, a plurality of moving grooves and placing grooves are formed in the top end and the interior of the kit body at equal intervals respectively, the moving grooves and the placing grooves are the same in number and correspond to each other in position in a one-to-one mode, and the moving grooves and the placing grooves correspond to each other in position in a one-to-one mode; two warning housings are symmetrically arranged in the moving groove; the warning housings are arranged on the two sides of the negative control tube, the positive control tube, the PCR reaction mixed liquid tube and the primer and probe mixed liquid tube, and when the negative control tube, the positive control tube, the PCR reaction mixed liquid tube or the primer and probe mixed liquid tube are damaged in a transportation diagram, the two warning housings can be closed due to weight loss, so that the warning effect is achieved, and the detection accuracy is improved. The problem that the negative control tube, the positive control tube, the PCR reaction mixed liquid tube or the primer and probe mixed liquid tube are not clearly damaged and are affected is avoided, and the use safety is improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of detection kits, and particularly relates to a detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well. Background Art

[0002] In the fields of life science and medicine, the research on the association between gene polymorphisms and human health and diseases has always been a hot topic. Among them, the polymorphisms of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene and the methionine synthase reductase (MTRR) gene occupy an important position in this research field.

[0003] The MTHFR gene is located at the position of chromosome lp36.3, with a full length of 19.3 kb and a total of 12 exons. The MTHFR encoded by it plays a key role in the folate-methionine cycle metabolic pathway, catalyzing the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, providing methyl raw materials for the synthesis of substances in the body and the methylation of nucleic acids and proteins, and at the same time maintaining the homocysteine in the body at a relatively low level. The common mutation site A1298C of the MTHFR gene will cause the encoded amino acid to change from glutamic acid to alanine, resulting in a decrease in enzyme activity, and further affecting folate metabolism.

[0004] The MTRR gene is also crucial in the folate metabolic pathway. The common mutation site A66G of the MTRR gene will affect the activity of methionine synthase, and further interfere with the metabolic process of homocysteine.

[0005] Numerous studies have confirmed that the polymorphisms of the MTHFR and MTRR genes are closely related to various diseases, including cardiovascular and cerebrovascular diseases and reproductive health, etc.

[0006] In terms of cardiovascular and cerebrovascular diseases, when the enzyme activity related to folate metabolism is low, even if folate is supplemented according to the normal amount, the folate level in the body may still be insufficient, resulting in the accumulation of homocysteine in the body, triggering hyperhomocysteinemia, and further increasing the incidence risk of cardiovascular and cerebrovascular diseases, such as systemic atherosclerosis and thrombosis, etc. A mild and moderate increase in homocysteine level can increase the risk of cardiovascular disease death by 4-6 times, and it is an independent risk factor for coronary heart disease.

[0007] In terms of reproductive health, the polymorphisms of the MTHFR and MTRR genes are closely related to diseases such as recurrent miscarriage and fetal neural tube defects. For example, the TT genotype of the C677T locus of the MTHFR gene is significantly correlated with low sperm concentration and high DNA fragmentation rate in male infertility patients, and may be a high-risk gene leading to male infertility.

[0008] In addition, these gene polymorphisms have also been found to be associated with the occurrence, development and prognosis of neurological diseases and tumors.

[0009] Currently, there are many methods for detecting MTHFR and MTRR gene polymorphisms, among which polymerase chain reaction-restriction fragment length polymorphism PCR-RFLP is more widely used.

[0010] Chinese patent application No. 202223533154.6 discloses a real-time fluorescence quantitative PCR detection kit for detecting human MTHFR gene, including a box body, a first box cover and a second box cover symmetrically arranged on the top of the box body, the first box cover and the second box cover are connected to the box body by sliding rods in a slide groove, and the first box cover and the second box cover are connected by a movable limit hook that can be slid out of or plugged into the limit groove. When in use, move the limit hook to disengage from the limit groove, push the first box cover and the second box cover in the opposite direction, and the sliders of the first box cover and the second box cover move to the maximum in the box body slide groove, so that the first box cover and the second box cover can be opened to the maximum extent.

[0011] If the internal test tube of the above-mentioned real-time fluorescence quantitative PCR human MTHFR gene detection kit is damaged during transportation, the tester will not be able to know it in advance during the test and may accidentally touch the damaged test tube and contact the reagent, which may easily affect it and reduce the safety of use.

[0012] In view of this, a detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well is designed to solve the above problems. Summary of the invention

[0013] To solve the problems raised in the above background technology, the present invention provides a detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well, which has the characteristics of improved safety and convenience, low detection cost and high detection throughput.

[0014] To achieve the above object, the present invention provides the following technical solution: A detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well, comprising: a box body, wherein a plurality of moving grooves and placing grooves are respectively arranged at equal intervals at the top end and inside of the box body, the number of moving grooves and placing grooves is the same and their positions correspond one by one, two warning cover shells are symmetrically arranged inside the moving grooves, a support plate is arranged inside the placing groove directly below the two warning cover shells, an elastic lifting mechanism is assembled between the support plate and the bottom end of the placing groove, a follow-up driving mechanism is assembled between the support plate and the two warning cover shells, two support rods are symmetrically arranged along the central axis at the top end of the support rod, the two support rods are located between the two warning cover shells and extend above the moving grooves, a placing ring with a rubber ring is fixedly connected to the top ends of the two support rods, a negative control tube and a positive control tube are respectively clamped inside the placing rings with rubber rings at one side edge, a plurality of PCR reaction mixture tubes are respectively clamped inside the middle placing rings with rubber rings, a plurality of primer and probe mixture tubes are respectively clamped inside the placing rings with rubber rings at the other side edge, a box cover is arranged outside the box body, a locking mechanism is assembled between the box cover and the box body, and pressing blocks are fixedly connected to the inner top wall of the box cover corresponding to the positions of the negative control tube, the positive control tube, the PCR reaction mixture tube and the primer and probe mixture tube.

[0015] Further, the elastic lifting mechanism includes two first springs symmetrically and fixedly connected to the bottom end of the placing groove, and the top ends of the first springs are fixedly connected to the bottom end of the support plate.

[0016] Further, when the negative control tube, the positive control tube, the PCR reaction mixture tube and the primer and probe mixture tube are in an unbroken state and are clamped inside the placing ring with a rubber ring, the support plate descends to the lowest point due to gravity, and the first spring is in a compressed state.

[0017] Further, the follow-up driving mechanism includes two first racks symmetrically and fixedly connected to the bottom end of the support plate, four driving shafts connected inside the placing groove through bearings, two first limiting rods symmetrically and fixedly connected inside the placing groove, and two moving frames fixedly connected to and symmetrically arranged on the two warning cover shells. The two first racks are located on the sides away from each other of the two first springs, the four driving shafts are symmetrically arranged in pairs on the sides away from each other of the two first racks, gears are fixedly sleeved on the driving shafts, the gears on the side close to the first rack are meshed and connected with the first rack, the gears on the same side are meshed and connected with each other, the two first limiting rods are located on the sides of the driving shafts on the same side away from the first rack and are perpendicular to the two first racks, the two moving frames are respectively sleeved on the two first limiting rods through holes, second racks are fixedly connected to the sides of the two moving frames close to each other, and the gears on the side close to the second rack are meshed and connected with the second rack.

[0018] Further, when the negative control tube, positive control tube, PCR reaction mixture tube and primer and probe mixture tube are damaged, the support plate rises to the highest point due to the loss of gravity, that is, when it fits with the bottom ends of the two warning covers, the two warning covers move to the closed state.

[0019] Further, when the two warning covers move to the closed state, the moving frame does not disengage from the first limiting rod.

[0020] Further, the number of the PCR reaction mixture tubes is the same as that of the primer and probe mixture tubes.

[0021] Further, the locking mechanism includes two second limiting rods symmetrically fixed on the inner wall of the box cover and four locking holes opened on the box cover. The second limiting rods are located above the box body. An activity frame is sleeved on the second limiting rods through holes. A second spring is elastically connected between the box cover and the activity frame outside the second limiting rods. Two locking blocks are symmetrically arranged on the side walls of the two activity frames away from each other. The four locking blocks correspond to the four locking holes in position and are respectively inserted into the four locking holes.

[0022] Further, when the locking block presses to open the box cover, the activity frame does not disengage from the second limiting rod.

[0023] Further, a positioning mechanism is assembled between the support plate and the placement groove. The positioning mechanism includes a support ring plate connected to the inside of the middle of the support plate through a bearing, two guide rods symmetrically fixed at the bottom end of the support plate, and a fixed seat fixed on the inner bottom wall of the placement groove. Two support rods are symmetrically fixed at the top end of the support ring plate. A screw rod is fixed at the bottom end of the support ring plate. A movable seat is connected to the screw rod through a transmission nut. The two guide rods are located outside the screw rod and inside the two first springs. The movable seat is sleeved on the two guide rods through holes. The position of the movable seat corresponds to the position of the fixed seat. Two magnets are symmetrically fixed inside the movable seat and the fixed seat respectively. The magnets on the movable seat and the magnets on the fixed seat have opposite polarities.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The present invention is provided with warning covers on both sides of the negative control tube, positive control tube, PCR reaction mixture tube and primer and probe mixture tube. When the negative control tube, positive control tube, PCR reaction mixture tube or primer and probe mixture tube is damaged during transportation, the two warning covers can be closed due to weightlessness, so as to play a warning role, avoid the problem that the detection personnel do not know the damage situation and contact the negative control tube, positive control tube, PCR reaction mixture tube or primer and probe mixture tube, and cause an impact on it, and improve the use safety.

[0025] 2. The present invention is provided with a positioning mechanism, which can position the support plate when removing the undamaged negative control tube, positive control tube, PCR reaction mixture tube, and primer and probe mixture tube, avoiding the closure of the two warning covers and improving the convenience of removal.

[0026] 3. The primer and probe mixture in the primer and probe mixture tube of the present invention can simultaneously detect the polymorphisms of human MTHFR and MTRR genes through a single reaction well, featuring low detection cost and high detection throughput. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 is a perspective view of the present invention; Figure 2 is a partial cross-sectional view of the present invention; Figure 3 is a partial longitudinal cross-sectional view of the present invention; Figure 4 is a schematic diagram of the elastic lifting mechanism and positioning mechanism of the present invention; Figure 5 is a schematic diagram of the follow-up drive mechanism of the present invention; Figure 6 of the present invention Figure 3 is an enlarged view of part A in; Figure 7 is a schematic diagram of the locking mechanism of the present invention; In the figure: 1, box body; 2, box cover; 3, negative control tube; 4, positive control tube; 5, PCR reaction mixture tube; 6, primer and probe mixture tube; 7, warning cover; 8, pressing block; 9, moving groove; 10, rubber ring placement ring; 11, support rod; 12, support plate; 13, placement groove; 101, first spring; 201, first rack; 202, gear; 203, drive shaft; 204, first limiting rod; 205, second rack; 206, moving frame; 301, movable frame; 302, locking block; 303, locking hole; 304, second spring; 305, second limiting rod; 401, support ring plate; 402, fixed seat; 403, magnet; 404, screw; 405, movable seat; 406, guide rod. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0028] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0029] Example 1 The present invention provides the following technical solution: A detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well, comprising: a box body 1, a plurality of moving grooves 9 and placing grooves 13 are respectively arranged at equal intervals at the top end and inside of the box body 1. The number of the moving grooves 9 and the placing grooves 13 is the same and their positions correspond one by one. Two warning covers 7 are symmetrically arranged inside the moving groove 9. A support plate 12 is arranged below the two warning covers 7 inside the placing groove 13. An elastic lifting mechanism is assembled between the support plate 12 and the bottom end of the placing groove 13. A follow-up driving mechanism is assembled between the support plate 12 and the two warning covers 7. Two support rods 11 are symmetrically arranged along the central axis at the top end of the support rod 11. The two support rods 11 are located between the two warning covers 7 and extend above the moving groove 9. A rubber ring-equipped placement ring 10 is fixedly connected to the top ends of the two support rods 11. A negative control tube 3 and a positive control tube 4 are respectively clamped inside the two rubber ring-equipped placement rings 10 at one side edge. A plurality of PCR reaction mixture tubes 5 are respectively clamped inside a plurality of rubber ring-equipped placement rings 10 in the middle. A plurality of primer and probe mixture tubes 6 are respectively clamped inside a plurality of rubber ring-equipped placement rings 10 at the other side edge. A box cover 2 is arranged outside the box body 1. A locking mechanism is assembled between the box cover 2 and the box body 1. Pressing blocks 8 are fixedly connected to the inner top wall of the box cover 2 corresponding to the positions of the negative control tube 3, the positive control tube 4, the PCR reaction mixture tubes 5 and the primer and probe mixture tubes 6.

[0030] In this embodiment, refer to the attached Figures 1-3, when using the detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well, release the locking state of the locking mechanism and lift the lower lid 2 of the extraction part. At this time, the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, and primer and probe mixture tube 6 lose the extrusion force and can be taken out. The unbroken negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, and primer and probe mixture tube 6 during transportation remain in place due to the gravity of the internal liquid and wait to be taken out. The broken negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, or primer and probe mixture tube 6 during transportation loses the gravity effect due to the loss of internal liquid. After the lid 2 is removed and the pressing force of the pressing block 8 is lost, the lower support plate 12 rises under the drive of the elastic lifting mechanism, driving the broken negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, or primer and probe mixture tube 6 and part of the structure of the follower drive mechanism to rise. The rise of part of the structure of the follower drive mechanism drives the other structures of the follower drive mechanism, driving the corresponding two warning covers 7 to move towards each other until the two warning covers 7 merge and stop. At this time, the broken negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, or primer and probe mixture tube 6 is covered by the corresponding two warning covers 7, which can remind the tester that the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, or primer and probe mixture tube 6 at this position is broken, avoiding the tester from contacting and being affected by it; The tester takes out the unbroken negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, and primer and probe mixture tube 6. First, open the tube caps of the negative control tube 3 and positive control tube 4, and pour the PCR reaction mixtures in the two PCR reaction mixture tubes 5 and the primer and probe mixtures in the two primer and probe mixture tubes 6 into the negative control tube 3 and positive control tube 4 respectively and mix evenly. After PCR amplification, fluorescence analysis is carried out to verify the effectiveness of the detection kit; After verifying the effectiveness of the detection kit, pour the PCR reaction mixture in the PCR reaction mixture tube 5 and the primer and probe mixture in the primer and probe mixture tube 6 into the test tube with the DNA sample and mix evenly. After PCR amplification, fluorescence analysis is carried out to judge the polymorphism situation of the MTHFR and MTRR genes in the DNA sample; The above PCR reaction mixture includes Taq DNA polymerase, dNTP mixture, 10×PCR buffer, MgCl2, UNG enzyme, and deionized water; In the primer and probe mixture, the primer and probe information designed for the A1298C locus of the MTHFR gene is as follows: Forward primer: 5'-CTGAAGGAGAAGGTGTCTGCGGGA-3' Reverse primer: 5'-AGGACGGTGCGGTGAGAGTG-3' Wild-type probe 1: FAM-5'-ACGCCCGGCTCCATG-3'-MGB Mutant probe 1: VIC-5'-ACGCCCGGCTCCCTG-3'-MGB Wild-type probe 2: ROX-5'-TACGCCCGGCTCCATG-3'-MGB Mutant probe 2: CY5-5'-TACGCCCGGCTCCCTG-3'-MGB The primer and probe information designed for the A66G site of the MTRR gene is as follows: Primer and probe information Forward primer: 5'-TGAAGGAGAAGGTGTCTGCGGGA-3' Reverse primer: 5'-AGGACGGTGCGGTGAGAGTG-3' Wild-type probe 1: FAM-5'-CCTGAAGCACGCGGCTG-3'-MGB Mutant probe 1: VIC-5'-CCTGAAGCACACGGCTG-3'-MGB Wild-type probe 2: ROX-5'-TCCTGAAGCACGCGGCTG-3'-MGB Mutant probe 2: CY5-5'-TCCTGAAGCACACGGCTG-3'-MGB.

[0031] Specifically, the elastic lifting mechanism includes two first springs 101 symmetrically fixed to the bottom end of the placement groove 13, and the top ends of the first springs 101 are fixed to the bottom end of the support plate 12.

[0032] In this embodiment, refer to the appendix Figure 4 , and the support plate 12 realizes lifting under the elastic force of the two first springs 101.

[0033] Specifically, when the negative control tube 3, the positive control tube 4, the PCR reaction mixture tube 5, and the primer and probe mixture tube 6 are in an intact state and clamped in the rubber-ringed placement ring 10, the support plate 12 descends to the lowest point due to gravity, and the first spring 101 is in a compressed state.

[0034] In this embodiment, refer to the appendix Figure 4, it can achieve that the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, and primer and probe mixture tube 6 that are not damaged during transportation remain in their original positions due to the gravitational force of the internal liquid and wait to be taken out, and the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, or primer and probe mixture tube 6 that are damaged during transportation lose their gravitational force due to the loss of internal liquid. After the lid 2 is removed and the pressing force of the pressing block 8 is lost, the lower support plate 12 rises under the drive of the elastic lifting mechanism.

[0035] Specifically, the follow-up drive mechanism includes two first racks 201 symmetrically fixed to the bottom end of the support plate 12, four drive shafts 203 connected to the inside of the placement groove 13 through bearings, two first limiting rods 204 symmetrically fixed to the inside of the placement groove 13, and two moving frames 206 fixed to and symmetrically arranged on the two warning covers 7. The two first racks 201 are located on the mutually remote sides of the two first springs 101. The four drive shafts 203 are arranged in pairs symmetrically on the mutually remote sides of the two first racks 201. A gear 202 is fixedly sleeved on the drive shaft 203. The gear 202 on the side close to the first rack 201 is meshed with the first rack 201. The two gears 202 on the same side are meshed with each other. The two first limiting rods 204 are located on the side of the same-side drive shaft 203 remote from the first rack 201 and are perpendicular to the two first racks 201. The two moving frames 206 are respectively sleeved on the two first limiting rods 204 through holes. Second racks 205 are respectively fixed to the mutually close sides of the two moving frames 206. The gear 202 on the side close to the second rack 205 is meshed with the second rack 205.

[0036] In this embodiment, referring to the attached Figure 3 and 5 , when the support plate 12 rises driven by the follow-up drive mechanism, the two first racks 201 rise. During the rising process of the two first racks 201, the two meshed gears 202 are driven to rotate. The two meshed gears 202 drive another two meshed gears 202 to rotate. The other two meshed gears 202 drive the two meshed second racks 205 to rotate. The two second racks 205 drive the two moving frames 206 to move towards each other on the two first limiting rods 204. The two moving frames 206 drive the two warning covers 7 to move towards each other to achieve the follow-up function of the two warning covers 7 and the support plate 12.

[0037] Specifically, when the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5, and primer and probe mixture tube 6 are in a damaged state, the support plate 12 rises to the highest point due to the loss of gravitational force, that is, when it fits with the bottom ends of the two warning covers 7, the two warning covers 7 move to the closed state.

[0038] In this embodiment, referring to the attached Figure 3 and5 It can realize the covering warning function for the negative control tube 3, positive control tube 4, PCR reaction mixture tube 5 or primer and probe mixture tube 6 damaged during transportation.

[0039] Specifically, when the two warning covers 7 move to the closed state, the moving frame 206 does not disengage from the first limiting rod 204.

[0040] In this embodiment, referring to the appendix Figure 5 to prevent the moving frame 206 from disengaging from the first limiting rod 204 and affecting its operation.

[0041] Specifically, the number of PCR reaction mixture tubes 5 is the same as the number of primer and probe mixture tubes 6.

[0042] In this embodiment, referring to the appendix Figure 1 to facilitate the detection work of the detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well.

[0043] Specifically, the locking mechanism includes two second limiting rods 305 symmetrically fixed to the inner wall of the box cover 2 and four locking holes 303 opened on the box cover 2. The second limiting rods 305 are located above the box body 1. An activity frame 301 is sleeved on the second limiting rods 305 through holes. A second spring 304 is elastically connected between the box cover 2 and the activity frame 301 outside the second limiting rods 305. Two locking blocks 302 are symmetrically arranged on the side walls of the two activity frames 301 away from each other. The four locking blocks 302 correspond to the four locking holes 303 one by one and are respectively inserted into the four locking holes 303.

[0044] In this embodiment, referring to the appendix Figure 3 , 6 and 7, the locking mechanism is to squeeze any one of the locking blocks 302 on both sides. The two locking blocks 302 being squeezed on both sides drive the two connected activity frames 301 to move towards each other on the two second limiting rods 305. The two activity frames 301 drive the other two locking blocks 302 to move in the same direction until the locking blocks 302 being squeezed on both sides disengage from the locking holes 303. At this time, the four locking blocks 302 are respectively disengaged from the four locking holes 303, and the separation of the box cover 2 and the box body 1 is completed.

[0045] Specifically, when the locking block 302 is squeezed to open the box cover 2, the activity frame 301 does not disengage from the second limiting rod 305.

[0046] In this embodiment, referring to the appendix Figure 7 to prevent the activity frame 301 from disengaging from the second limiting rod 305 and affecting its operation.

[0047] Embodiment 2 The difference between this embodiment and Embodiment 1 is: Specifically, a positioning mechanism is assembled between the support plate 12 and the placement groove 13. The positioning mechanism includes a support ring plate 401 connected to the inside of the exact middle of the support plate 12 through a bearing, two guide rods 406 symmetrically fixed to the bottom end of the support plate 12, and a fixed seat 402 fixed to the inner bottom wall of the placement groove 13. Two support rods 11 are symmetrically fixed to the top end of the support ring plate 401. A screw rod 404 is fixed to the bottom end of the support ring plate 401. A movable seat 405 is connected to the screw rod 404 through a transmission nut. The two guide rods 406 are located outside the screw rod 404 and inside two first springs 101. The movable seat 405 is sleeved on the two guide rods 406 through an opening. The position of the movable seat 405 corresponds to the position of the fixed seat 402. Two magnets 403 are symmetrically fixed to the inside of the movable seat 405 and the fixed seat 402 respectively. The magnet 403 on the movable seat 405 and the magnet 403 on the fixed seat 402 have opposite polarities.

[0048] In this embodiment, refer to the attached Figure 3 and 4 , the negative control tubes 3, positive control tubes 4, PCR reaction mixture tubes 5, and primer and probe mixture tubes 6 that were not damaged during transportation remain in place due to the gravity of the internal liquid and await removal. Before removal, the undamaged negative control tubes 3, positive control tubes 4, PCR reaction mixture tubes 5, or primer and probe mixture tubes 6 are rotated. The undamaged negative control tubes 3, positive control tubes 4, PCR reaction mixture tubes 5, or primer and probe mixture tubes 6 drive the rubber-ringed placement ring 10 to rotate (the clamping force between the rubber-ringed placement ring 10 and the placement tube is set to be able to drive the rotation). The rubber-ringed placement ring 10 drives the two support rods 11 to rotate. The two support rods 11 drive the support ring plate 401 to rotate. The support ring plate 401 drives the screw rod 404 to rotate. During the rotation of the screw rod 404, the movable seat 405 moves up and down on the screw rod 404 under the guiding action of the two guide rods 406 until it stops rotating. At this time, the movable seat 405 fits with the fixed seat 402, and the upper and lower magnets 403 adsorb and fix the position of the support plate 12. That is, during the removal of the undamaged negative control tubes 3, positive control tubes 4, PCR reaction mixture tubes 5, or primer and probe mixture tubes 6, the corresponding two warning covers 7 will not close, and it does not affect the removal.

[0049] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well, characterized in that, Comprising: A box body (1), on the top end and inside of which a plurality of moving grooves (9) and placing grooves (13) are respectively arranged at equal intervals. The number of the moving grooves (9) and the placing grooves (13) is the same and their positions correspond one by one. Two warning covers (7) are symmetrically arranged inside the moving groove (9). A support plate (12) is arranged inside the placing groove (13) directly below the two warning covers (7). An elastic lifting mechanism is assembled between the support plate (12) and the bottom end of the placing groove (13). A follow-up driving mechanism is assembled between the support plate (12) and the two warning covers (7). At the top end of the support rod (11), two support rods (11) are symmetrically arranged along the central axis. The two support rods (11) are located between the two warning covers (7) and extend above the moving groove (9). At the top ends of the two support rods (11), a rubber ring-equipped placing ring (10) is fixedly connected. Inside the two rubber ring-equipped placing rings (10) at one side edge, a negative control tube (3) and a positive control tube (4) are respectively clamped tightly. Inside a plurality of the rubber ring-equipped placing rings (10) in the middle, a plurality of PCR reaction mixture tubes (5) are respectively clamped tightly. Inside a plurality of the rubber ring-equipped placing rings (10) at the other side edge, a plurality of primer and probe mixture tubes (6) are respectively clamped tightly. A box cover (2) is arranged outside the box body (1). A locking mechanism is assembled between the box cover (2) and the box body (1). On the inner top wall of the box cover (2), pressing blocks (8) are fixedly connected corresponding to the positions of the negative control tube (3), the positive control tube (4), the PCR reaction mixture tubes (5) and the primer and probe mixture tubes (6).

2. The detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well according to claim 1, wherein: The elastic lifting mechanism includes two first springs (101) symmetrically and fixedly connected to the bottom end of the placing groove (13), and the top ends of the first springs (101) are fixedly connected to the bottom end of the support plate (12).

3. The detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well according to claim 2, wherein: When the negative control tube (3), the positive control tube (4), the PCR reaction mixture tubes (5) and the primer and probe mixture tubes (6) are in an intact state and are clamped tightly in the rubber ring-equipped placing ring (10), the support plate (12) descends to the lowest point due to gravity, and the first spring (101) is in a compressed state.

4. A detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 3, characterized in that: The follow-up driving mechanism includes two first racks (201) symmetrically and fixedly connected to the bottom end of the support plate (12), four driving shafts (203) connected to the inside of the placement groove (13) through bearings, two first limiting rods (204) symmetrically and fixedly connected to the inside of the placement groove (13), and two moving frames (206) fixedly connected to and symmetrically arranged on the two warning covers (7). The two first racks (201) are located on the sides away from each other of the two first springs (101). The four driving shafts (203) are symmetrically arranged in pairs on the sides away from each other of the two first racks (201). A gear (202) is fixedly sleeved on the driving shaft (203). The gear (202) close to the first rack (201) is meshed and connected with the first rack (201). The gears (202) on the same side are meshed and connected. The two first limiting rods (204) are located on the sides of the driving shafts (203) on the same side away from the first racks (201) and are perpendicular to the two first racks (201). The two moving frames (206) are respectively sleeved on the two first limiting rods (204) through openings. Second racks (205) are respectively fixedly connected to the sides of the two moving frames (206) close to each other. The gear (202) close to the second rack (205) is meshed and connected with the second rack (205).

5. The detection kit for simultaneously detecting the polymorphisms of human MTHFR and MTRR genes in a single reaction well according to claim 4, characterized in that: When the negative control tube (3), the positive control tube (4), the PCR reaction mixture tube (5), and the primer and probe mixture tube (6) are in a damaged state, the support plate (12) rises to the highest point due to the loss of gravity, that is, when it fits against the bottom ends of the two warning covers (7), the two warning covers (7) move to a closed state.

6. The detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 5, characterized in that: When the two warning covers (7) move to a closed state, the moving frames (206) do not disengage from the first limiting rods (204).

7. A detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 6, characterized in that: The number of the PCR reaction mixture tubes (5) is the same as the number of the primer and probe mixture tubes (6).

8. The detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 7, wherein: The locking mechanism includes two second limiting rods (305) symmetrically and fixedly connected to the inner wall of the box cover (2) and four locking holes (303) opened on the box cover (2). The second limiting rods (305) are located above the box body (1). An activity frame (301) is sleeved on the second limiting rods (305) through openings. A second spring (304) is elastically connected between the outside of the second limiting rods (305) and between the box cover (2) and the activity frame (301). Two locking blocks (302) are symmetrically arranged on the side walls of the two activity frames (301) away from each other. The four locking blocks (302) correspond to the four locking holes (303) in position and are respectively inserted into the four locking holes (303).

9. A detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 8, characterized in that: When the locking blocks (302) are used to squeeze and open the box cover (2), the activity frame (301) does not disengage from the second limiting rods (305).

10. A detection kit for simultaneously detecting human MTHFR and MTRR gene polymorphisms in a single reaction well according to claim 9, characterized in that: A positioning mechanism is assembled between the support plate (12) and the placement groove (13). The positioning mechanism includes a support ring plate (401) connected to the inside of the exact middle of the support plate (12) through a bearing, two guide rods (406) symmetrically fixed to the bottom end of the support plate (12), and a fixed seat (402) fixed to the inner bottom wall of the placement groove (13). Two support rods (11) are symmetrically fixed to the top end of the support ring plate (401). A screw rod (404) is fixed to the bottom end of the support ring plate (401). A movable seat (405) is connected to the screw rod (404) through a transmission nut. The two guide rods (406) are located outside the screw rod (404) and inside two first springs (101). The movable seat (405) is sleeved on the two guide rods (406) through an opening. The position of the movable seat (405) corresponds to the position of the fixed seat (402). Two magnets (403) are symmetrically fixed to the inside of the movable seat (405) and the fixed seat (402) respectively. The magnets (403) on the movable seat (405) and the magnets (403) on the fixed seat (402) have opposite polarities.

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