Integration reagent tube for luminescence immunodetection, and use method thereof

[0035] Example one (The bottom surface of the microporous cup is integrally formed with a ball as a reflector).

[0036] Such as Figure 1 ~ Figure 5 The shown integrated reagent tube for luminescence immunoassay includes a microporous cup 10 that can be solid-phased in advance and a sample filter device 20 for filtering the liquid to be tested.

[0037] Such as figure 1 As shown, the microporous cavity of the microporous cup 10 of the present invention is composed of a circular through hole 11 in the upper part and a cup hole 12 in the lower part having a side ring wall 121 that increases the area of ​​upward projection and reflection. The hole is a rounded table hole cup hole 12, the bottom surface 122 of the cup hole 12 is protrudingly provided with a reflector 30 to increase the area of ​​upward projection and reflection, and a ball 31 is used as a reflector (30 is integrally formed and fixed in the cup hole On the bottom surface 122 of 12, the sphere 31 can reflect light to the side ring surface 121 of the cup hole 12; the solid-liquid reaction interface between the side ring wall 121 of the cup hole 12 of the present invention and the detection liquid of the object to be measured increases, so It is conducive to the full progress of immune reaction, luminescence reaction or enzyme-catalyzed reaction, and the time for the reaction to reach equilibrium is greatly shortened, thereby improving detection sensitivity and detection efficiency; the inclined structure of the solid-liquid reaction interface between the ring wall 121 on the side of the cup hole 12 and the detection liquid The generated luminous light effectively converges, so that the photomultiplier tube (PMT) probe of the detection instrument receives more light signals at the solid-liquid interface to improve the detection sensitivity.

[0038] Such as figure 1 , figure 2 , Figure 4 with Figure 5 As shown, the sample filtering device 20 of the present invention includes a sleeve 21 that is detachably fitted with the mouth of the microporous cup 10, and a plastic sample filter cup 22 fixed on the sleeve 21 is suspended on the microporous cup 10. In the upper half of the inner part, the shape of the sleeve 21 is a ring that fits with the mouth of the microporous cup 10, that is, the outer wall of the circular through hole 11. The sample filter cup 22 is arranged at the center of the sleeve 21, and the sample filter cup 22 The outer wall and the sleeve 21 are connected and fixed by a plurality of spacer beams 211 spaced apart. The space 212 of the adjacent spacer beams 211 is connected to the inside and outside of the mouth of the microporous cup 10; the bottom surface 222 of the cup cavity 221 of the sample filter cup 22 of the present invention A binding membrane 23 formed by stacking a sample filter membrane 231 and a marker film 232 from top to bottom is fixed on the top. The sample filter device 20 also includes a snap ring 24 that presses and fixes the edge of the binding membrane 23 and is sleeved into the sample filter cup The snap ring 24 in the cup cavity 221 presses the bonding film 23 on the bottom surface 222 of the cup cavity 221; the bottom surface 222 of the sample filter cup 22 is provided with a drip tube 223 connecting the inside and outside of the cup cavity 221 of the sample filter cup 22.

[0039] Such as Figure 3 ~ Figure 5 As shown, the sample filter device 20 is sleeved on the mouth of the microporous cup 10 to form the integrated reagent tube for luminescence immunoassay of the present invention.

[0040] The method of using the integrated reagent tube for luminescence immunoassay of the present invention will be described in detail below.

[0041] The first step: such as figure 2 As shown, the microporous cavity of the microporous cup 10 is solid-phased to match the marker film 232 in the sample filter cup 22;

[0042] The second step: such as Figure 3 ~ Figure 5 As shown, the sample filter device 20 is fixed at the mouth of the microporous cup 10; a specified liquid volume of the analyte sample is added to the sample filter cup 22, and the cell particles in the analyte are blocked by the sample filter membrane 231 on the upper layer of the membrane. The water phase infiltrates down from the pores of the sample filter membrane 231 to the marker film 232. The marker in the marker film 232 is dissolved by the water phase and initially combined with the test substance in the water phase in the marker film 232 to obtain the initial Reactant;

[0043] The third step: such as Figure 4 , Figure 5 As shown, the pipe joint of the air pressure pump is sleeved on the outer wall of the top end of the sample filter cup 22 through the inclined ring surface, and the initial reactant in the sample filter cup 22 flows into the inner wall of the microporous cup 10 (cup hole 12, the solid phase on the side wall 121, the bottom wall 122 and the surface of the spherical cavity 31), even if the non-cellular components and markers in the initial reactant enter the solid phase area in the microporous cup 10 to participate in the reaction; Phase-to-be-detected molecule-label reaction chain to obtain the target for luminescence immunoassay;

[0044] The fourth step: such as figure 2 As shown, the sample filter device 20 is pulled out from the microporous cup 10 and discarded;

[0045] Step 5: Wash the microporous cavity of the microporous cup 10, continuously inject the lotion from the opening of the microporous cavity, and use a negative pressure device to continuously suck and wash thoroughly so that unreacted components are sucked out of the micropores Cup 10

[0046] Step 6: After washing the microporous cup 10, add a certain volume of luminescent substrate liquid to the microporous cup 10 to form a luminescence reaction, measure the luminous intensity within a certain period of time, and calculate the corresponding to-be-tested from the stored calibration curve Concentration and report test results.

[0047] In the third step of the above method of use, negative pressure can also be used to make the liquid in the sample filter cup 22 inject into the microporous cup 10 through the dropper 223; in the sixth step, because of the cup hole side ring at the bottom of the microporous cavity The inclined surface structure of the solid-liquid reaction interface between the wall and the detection liquid can effectively converge the generated luminous light, so that the photomultiplier tube (PMT) probe of the detection instrument can receive more light signals from the solid-liquid interface, which can improve the detection sensitivity .

[0048] Example two (The reflector integrally formed on the bottom of the microporous cup can be replaced with other structures).

[0049] Such as Image 6 The shown integrated reagent tube for luminescence immunoassay is different from the first example in that the ball gap 31 in the first embodiment is replaced with one of a cone 32, a truncated cone 33, a pyramid 34 or a cone 35; 32. The truncated cone 33, the pyramid 34 or the prism 35 can increase the solid-liquid interface in the microporous cup 10, and the light on it can be reflected to the side wall of the cup hole 12 of the microporous cup 10 and concentrated during light detection. The photomultiplier tube (PMT) probe of the detection instrument can receive more light signals at the solid-liquid interface; the sample filter device 20 of the integrated reagent tube for luminescence immunoassay in this embodiment is the same as that in the first embodiment. The method of using the integrated reagent tube for detection is the same as in Example 1.

[0050] Example three (A number of tetrahedrons are integrally formed on the side ring wall 121 of the cup hole 12 of the microporous cup).

[0051] Such as Figure 7 As shown, the sample filtering device 20 of this embodiment is the same as that of Embodiment 1. The ring wall 121 on the side of the cup hole 12 of the microporous cup of this embodiment is integrally formed with several tetrahedrons 36. These tetrahedrons 36 increase the solid-liquid reaction. The interface, and the solid-liquid reaction interface can be effectively irradiated by the projected light and reflected and converged, so that the light signal detected by the photomultiplier tube (PMT) probe of the detection instrument is increased, and the detection sensitivity is greatly improved; the integrated reagent tube of this embodiment The method of use is the same as in Example 1.