WO2016026165A1

WO2016026165A1 - Trace sample test paper

Info

Publication number: WO2016026165A1
Application number: PCT/CN2014/085711
Authority: WO
Inventors: 岳朋; 周锦源; 盛威玮
Original assignee: 深圳市领治医学科技有限公司
Priority date: 2014-08-18
Filing date: 2014-09-02
Publication date: 2016-02-25
Also published as: CN104142392B; CN104142392A

Abstract

Disclosed is a trace sample test paper, comprising a test paper body (20) and a sample collection device (10). A blood cell membrane (26) is provided on the inner wall of a negative pressure compartment (27) of the test paper body (20) above a sample pad (21) of the test paper body (20), wherein the blood cell membrane (26) is provided with a stabilizing solution reservoir tank (28), and the sample collection device (10) comprises a negative pressure element (11), and an anticoagulant sample tube (12). The anticoagulant sample tube (12) set in the sample collection device (10) can significantly reduce the blood clotting phenomenon of samples, reduce the loss of blood plasma, and thereby reduce the collection amount of blood samples. The blood samples are dropped into the stabilizing solution reservoir tank (28), and this can stabilize and dilute blood samples, further reducing the blood clotting phenomenon, so that the blood plasma in the blood samples can be more fully utilized, and thus the fingertip blood can be used as blood samples in rapid tests, and the collection amount of blood samples can be less than 1 μL. The negative pressure compartment (27) makes the liquid in the stabilizing solution reservoir tank (28) flow more fully and quickly into the sample pad (21) and thereby improves the utilization rate of blood samples.

Description

Micro sample test strip

Technical field

The invention relates to the field of in vitro detection technology, in particular to a micro sample detection test paper.

Background technique

With the advancement of society and the improvement of living standards, people are paying more and more attention to their own health, and more and more attention is paid to the early diagnosis and screening of diseases. Therefore, more accurate and convenient diagnostic tests are needed to meet this requirement. Diagnostic test strips, as a widely used testing method widely used by researchers, medical workers and users, are characterized by fast, convenient and low price, and are receiving more and more attention from everyone.

Rapid diagnostic test strips are a solid reagent developed over the past two decades. It has the advantages of simplicity, speed, accuracy and low cost. The easiest way is to use a test strip, dip the specimen, and compare it with the standard swatch to get the test results. Because it has got rid of valuable equipment and equipment, it has greatly reduced the cost of testing, and it has been welcomed by the majority of medical and health units. It is of great significance to the medical inspection of the vast rural and industrial and mining units in China.

China has developed rapidly in the rapid diagnosis of test strips, and the varieties are no less than the United States and Europe. For example, drug-sensitive test strips, urea nitrogen quantitative test strips, etc. have been used for decades; a number of joint test strips (such as Balian, Qilian, Liulian, etc.) have also been successfully developed. New progress has been made in the sensitivity and shelf life of test strips, and individual projects have reached international advanced levels. However, the most difficult problem for the current rapid diagnostic test strips is that the sample collection required for the test strip is large, especially in the blood. Because the fingertip blood is extremely clotting, it is impossible to apply the fingertip blood sample for detection, and it is necessary to collect venous blood, and Still need at least 20ul The sample size, therefore the collection of blood samples requires professional operations, it is difficult to spread to the hands of the majority of users.

Summary of the invention

The invention aims at the problem that the existing rapid test test paper cannot apply the fingertip blood sample, and the required sample amount is large, which leads to inconvenient sample collection, and provides a fingertip blood test sample, and only a small sample can be used. A small sample test strip for testing is performed.

In order to achieve the above object, the present invention adopts the following technical solutions.

A micro-sample test strip comprising a test strip body and a sample collection device, the test strip body comprising a bottom plate, and a sample pad, a bonding pad, a reaction film and an absorption pad disposed on the bottom plate and closely connected in sequence, the reaction A detection zone and a quality control zone are provided on the membrane.

The sample pad is provided with a blood cell filter membrane, and the blood cell filter membrane is provided with a stable liquid storage tank, the groove surface of the stable liquid storage tank is provided with a tank cover, and the bottom of the stable liquid storage tank is provided There is a first-class liquid port, and a liquid port is provided on the liquid port.

Further, the slot cover is a slot mask, and the slot mask is provided with a slot mask to expose the ear; the sealing member is a mouth film, and the sealing film is sealed around the flow port around the nozzle. A sealing film extending to the outside of the stable liquid storage tank is provided on the sealing film to expose the ear.

Preferably, the stabilizing liquid storage tank is provided with a blood stabilizing liquid, and the blood stabilizing liquid is an EDTA buffer having a pH of 6.8-7.6.

The sample collector includes a negative pressure member and an anticoagulation sample tube, and one end of the anticoagulation sample tube is in communication with the negative pressure member.

Further, the anticoagulation sample tube is a siphon tube containing an anticoagulant, one end of the siphon tube is connected to the negative pressure member, and the other end of the siphon tube is provided with a tube film.

Preferably, the anticoagulant is from 0.1 to 1.5% by weight of a heparin solution.

The test paper body further includes a negative pressure chamber, the bottom plate, the sample pad, the bonding pad, the reaction film, the absorption pad and the blood cell filter are disposed in the negative pressure chamber, and the stable liquid storage tank is disposed on the negative pressure chamber, and is stable The liquid storage tank and the negative pressure chamber can be connected through the liquid discharge port.

Further, a gap is provided between the sample pad and the blood cell filter.

Further, the pressure of the negative pressure chamber is 10-300 Pa.

Compared with the prior art, the invention has the beneficial effects that the invention can significantly reduce the coagulation phenomenon of the blood sample and reduce the loss of plasma by setting the anticoagulation sample tube in the sample collector, thereby reducing the collection amount of the blood sample. . A stabilizing liquid storage tank is arranged on the sample pad, and the blood sample is added to the stable liquid storage tank to stabilize and dilute the blood sample, thereby further reducing the blood coagulation phenomenon, thereby making the blood plasma in the blood sample more fully utilized, and making the fingertip Blood can be used as a test sample for rapid test strips. By setting the negative pressure chamber, the liquid in the liquid storage tank can be more completely and quickly flowed into the sample pad, thereby improving the utilization rate of the sample. The pressure of the negative pressure chamber is 10-300Pa, so that the liquid can quickly reach the sample pad from the liquid storage tank, and the blood cell breakage caused by excessive pressure can be avoided, and the final detection result is disturbed. The sample collector is provided with a siphon tube containing an anticoagulant, and the anticoagulant in the siphon tube is extruded before the blood sample is collected, and an anticoagulant film is formed on the inner wall of the siphon tube, and the anticoagulant film is formed by the anticoagulant film. It has anticoagulant effect on blood, and the setting of the sample collector is simple and easy to operate. By setting the sample collector, the liquid storage tank and the negative pressure chamber, the invention can accurately detect the disease by collecting only a small amount of fingertip blood samples, and the sample collection amount can be less than 1 μL.

DRAWINGS

1 is a schematic view showing the structure of the sample collector of Embodiment 1 when an anticoagulant is not loaded;

2 is a schematic view showing the structure of the sample collector of Embodiment 1 with an anticoagulant;

3 is a schematic structural view of the anticoagulant in the sample collector of FIG. 2 after extrusion;

4 is a schematic structural view of the sample pad, the bonding pad, the reaction film, and the absorption pad of Example 1 after being disposed on the bottom plate;

Figure 5 is a perspective view showing the structure of the test paper body of the first embodiment;

Figure 6 is a schematic structural view of the A-A cross section of Figure 5;

Figure 7 is a plan view of the test paper body of the first embodiment;

Figure 8 is a plan view showing the test paper body of the first embodiment after the groove mask is removed;

Fig. 9 is a schematic view showing the structure of the test paper body of the first embodiment after being loaded into a plastic support frame.

detailed description

In order to more fully understand the technical content of the present invention, the technical solutions of the present invention will be further described and illustrated below in conjunction with specific embodiments.

Example 1

1-9, a rapid test strip for diagnosing heart disease provided by the present embodiment includes a test strip body 20, a sample collector 10, and a plastic support frame 30. Wherein, the test paper body 10 is composed of a bottom plate 25, a sample pad 21, a bonding pad 22, a reaction film 23, an absorption pad 24, a blood cell filter 26, a stabilizing liquid storage tank 28, and a negative pressure chamber 27; the sample collector 10 is under negative pressure. The piece 11 and the anticoagulation sample tube 12 are constructed.

As shown in FIG. 1-2, the negative pressure member 11 in the sample collector 10 is an elastic ball, and the anticoagulation sample tube 12 is a siphon tube containing an anticoagulant 14, and one end of the siphon tube is sealed and inserted into the elastic ball. The siphon is connected to the elastic ball, and the other end of the siphon is sealed with a sealing membrane 13 to keep the anticoagulant 14 in the siphon. In this embodiment, the anticoagulant 14 in the siphon tube is a 0.3 wt% heparin solution. Before use, the sealing film 13 is removed, and the anticoagulant in the siphon is squeezed by pressing the elastic ball. 14 is discharged to form an anticoagulant film 15 on the inner wall of the siphon, as shown in FIG. The siphon is then moved to the sample to aspirate the blood sample, which has a layer of anticoagulant 15 in the siphon that significantly reduces blood clotting.

As shown in FIG. 4, the sample pad 21, the bonding pad 22, the reaction film 23, and the absorption pad 24 are respectively disposed on the bottom plate 25 and are closely connected in sequence; meanwhile, a detection area 231 and a quality control area are disposed in parallel on the reaction film 23. 232, and the detection zone 231 is located between the quality control zone 232 and the bonding pad 22 to form a test paper substrate. The detection zone 231, that is, the reaction membrane 23 is coated with an antibody capable of specifically binding to the antigen to be tested, and in this embodiment, the detection zone 231 is coated with a troponin I antibody; The region 232, that is, the reaction membrane 23 is coated with an antigen capable of specifically binding to the tracer. In the present embodiment, the 232 region is coated with the troponin I antigen. The bond pad 22 is comprised of a block of glass fibers and a colloidal gold tracer coated on a block of glass fibers. (In other embodiments, other substances may also be coated in the bond pad 22, the detection zone 231, and the quality control zone 232 based on disease detection principles and existing research findings.)

As shown in FIG. 5 and FIG. 6, the test paper substrate is placed in the negative pressure chamber 27, and a blood cell filter membrane 26 is disposed on the inner wall of the negative pressure chamber 27 above the sample pad 21 to make the blood cell filter membrane 26 and the sample pad 21 There is a gap between them, and the pressure in the negative pressure chamber 27 is set to 30 Pa by evacuation. A stabilizing liquid storage tank 28 is disposed above the negative pressure chamber 27 and the blood cell filter membrane 26. The bottom of the tank of the stabilized liquid storage tank 28 is provided with a first-rate liquid port through which the negative pressure chamber 27 and the stabilizing liquid can be The reservoir 28 is in communication. A mouthpiece is arranged on the liquid discharge port. The sealing member in the embodiment is a mouth film 284. The sealing film 284 is sealed around the liquid flow port, and the sealing film 284 is provided with a solution extending to the stable liquid reservoir. The outer sealing film of 28 is 283. The liquid discharge port can be temporarily sealed by the sealing film 284, so that the negative pressure chamber 27 becomes a closed space. When the negative pressure chamber 27 and the stabilizing liquid storage tank 28 need to be connected, the passage can be passed. The sealing film 284 is peeled off by the lifting sealing film 283. A groove cover is disposed on the groove surface of the stabilization liquid storage tank 28. The groove cover in the embodiment is a groove surface film 281, and the circumference of the groove surface film 281 is sealed and pasted with the groove wall of the stable liquid storage tank 28 to make a stable liquid. The reservoir 28 becomes a closed space. At the same time, a trough mask 282 is further disposed on the trough mask 281. As shown in FIG. 5 and FIG. 7-8, when the trough mask 281 of the stabilizing liquid reservoir 28 needs to be opened, the trough 282 is removed through the trough mask. can. The blood stasis liquid is placed in the stabilizing liquid reservoir 28 to form the test paper body 20. The blood stabilizing solution in this example was an EDTA buffer having a pH of 7.0.

As shown in FIG. 9, the test paper body 20 is loaded into a plastic support frame 30. The plastic support frame 30 is provided with an injection window 32 and an observation window 31. The sample window 32 of the plastic support frame 30 can drop the sample. It is added to the stabilizing liquid reservoir 28, and the change of the detecting zone 231 and the quality control zone 232 can be observed through the observation window 31. The prepared rapid test test paper was recorded as TP1.

In other embodiments, other anticoagulant substances may also be used as the anticoagulant 14 in the siphon tube, and when the heparin solution is used as the anticoagulant 14, 0.1-1.5 wt% may also be used. Heparin solution. The blood stabilizing solution in the stabilizing solution reservoir 28 may also be an EDTA buffer having a pH of 6.8-7.6.

The method for using the micro sample detection test paper: firstly tearing off the sealing film 13 at one end of the siphon tube on the sample collector 10, squeezing the negative pressure member 11 to exclude the anticoagulant 14 in the siphon tube, and then moving the siphon tube to the fingertip The blood is shed and the negative pressure member 11 is released, so that the fingertip blood enters the siphon tube under the negative pressure of the sample collector 10, and becomes a blood sample to be detected. Then, the sample cover 10 is used to pierce the cover film 281 on the stable liquid storage tank 28 through the injection window 32, and the blood sample in the siphon tube is squeezed into the stable liquid storage tank 28, and then the left and right sides are slightly shaken to stabilize the liquid storage. The liquid tank 28 uniformly dilutes the blood sample in the blood stabilizing liquid. Next, the cover mask 281 is removed by first pulling the cover mask 282, The sealing film 284 is removed and the sealing film 284 is taken out to connect the negative pressure chamber 27 with the stabilizing liquid storage tank 28. The blood sample diluent in the stabilizing solution reservoir 28 rapidly flows through the blood cell membrane 26 under the dual action of gravity and negative pressure and enters the sample pad 21, wherein the blood cells are trapped on the blood cell membrane 26. Under the action of capillary siphon, the liquid sequentially flows through the sample pad 21, the bonding pad 22, the detection zone 231, the quality control zone 232, and the absorption pad 24 and performs a corresponding specific reaction. Finally, the inspection result is observed through the observation window 31.

Example 2

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 10 Pa. The prepared rapid test test paper was recorded as TP2.

Example 3

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 20 Pa. The prepared rapid test test paper was recorded as TP3.

Example 4

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 100 Pa. The prepared rapid test test paper was recorded as TP4.

Example 5

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 200 Pa. The prepared rapid test test paper was recorded as TP5.

Example 6

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 300 Pa. The prepared rapid test test paper was recorded as TP6.

Example 7

The rapid test strip for detecting heart disease provided by the embodiment has the same structure as the quick test strip described in Embodiment 1, except that the pressure in the negative pressure chamber is set to 400 Pa. The prepared rapid test test paper was recorded as TP7.

Comparative example 1

According to the prior art, a rapid test strip for detecting heart disease is prepared. Specifically, the rapid test strip includes a bottom plate, and the sample pad, the bonding pad, the reaction film and the absorption pad are respectively disposed on the bottom plate and are closely connected in sequence. At the same time, a detection zone and a quality control zone are arranged in parallel on the reaction film, and the detection zone is located between the quality control zone and the bonding pad. The detection zone is coated with troponin I antibody, the control region is coated with troponin I antigen, and the binding pad is composed of a glass fiber block and a colloidal gold tracer coated on the glass fiber block. (The material coated in the bonding pad, the detection zone and the quality control zone is exactly the same as in Example 1.) Then, it is packaged in a plastic support frame, and the sample can be dropped onto the sample pad through the injection window of the plastic support frame, and observed. The window can observe changes in the detection zone and the quality control zone. The prepared rapid test test paper was recorded as TP8.

The fingertip blood of 100 patients with heart disease was collected according to the collected amounts of 0.8 μL, 1 μL, 2 μL, 10 μL, and 20 μL, respectively, and detected by the rapid test strips TP1-TP8 prepared in the above examples and comparative examples. The detection rates of each rapid test strip are shown in the table below.

	0.8μL0.8μL	1μL1μL	2μL2μL	10μL10μL	20μL20μL
	0.8μL0.8μL	1μL1μL	2μL2μL	10μL10μL	20μL20μL	TP 1TP 1	100％100%	100％100%	100％100%	100％100%	100％100%

TP 2TP 2	100％100%	100％100%	100％100%	100％100%	100％100%
TP 2TP 2	100％100%	100％100%	100％100%	100％100%	100％100%	TP 3TP 3	100％100%	100％100%	100％100%	100％100%	100％100%
TP 4TP 4	50％50%	60％60%	65％65%	70％70%	75％75%	TP 3TP 3	100％100%	100％100%	100％100%	100％100%	100％100%
TP 4TP 4	50％50%	60％60%	65％65%	70％70%	75％75%	TP 5TP 5	30％30%	40％40%	45％45%	55％55%	60％60%
TP 6TP 6	10％10%	20％20%	20％20%	25％25%	30％30%	TP 5TP 5	30％30%	40％40%	45％45%	55％55%	60％60%
TP 6TP 6	10％10%	20％20%	20％20%	25％25%	30％30%	TP 7TP 7	5％5%	7％7%	10％10%	15％15%	20％20%
TP 8TP 8	00	00	00	00	20％20%	TP 7TP 7	5％5%	7％7%	10％10%	15％15%	20％20%

It can be seen from the data in the above table that the present invention can accurately detect diseases by collecting a small sample by setting a collecting device and providing a negative pressure chamber and a stable liquid storage tank on the sample pad, and the sample collection amount can be less than 1 μL.

The above description is only for the purpose of further illustrating the technical content of the present invention in order to facilitate the understanding of the reader, but the embodiment of the present invention is not limited thereto, and any technology extension or re-creation according to the present invention is subject to Protection of the invention.

Claims

A micro-sample test strip comprising a test strip body and a sample collection device, the test strip body comprising a bottom plate, and a sample pad, a bonding pad, a reaction film and an absorption pad disposed on the bottom plate and closely connected in sequence, the reaction The membrane is provided with a detection zone and a quality control zone, which are characterized by:

The sample pad is provided with a blood cell filter membrane, and the blood cell filter membrane is provided with a stable liquid storage tank, the groove surface of the stable liquid storage tank is provided with a tank cover, and the bottom of the stable liquid storage tank is provided There is a first-class liquid port, and a liquid port is provided on the liquid port;

The sample collector includes a negative pressure member and an anticoagulation sample tube, and one end of the anticoagulation sample tube is in communication with the negative pressure member.
A micro sample detecting test paper according to claim 1, wherein the test paper body further comprises a negative pressure chamber, and the bottom plate, the sample pad, the bonding pad, the reaction film, the absorption pad and the blood cell filter are set at a negative In the pressure chamber, the stable liquid storage tank is arranged on the negative pressure chamber, and the stable liquid storage tank and the negative pressure chamber can be connected through the liquid discharge port.
A micro sample detecting test paper according to claim 2, wherein a gap is provided between the sample pad and the blood cell filter.
A micro sample detecting test paper according to claim 3, wherein the negative pressure chamber has a pressure of 10 to 300 Pa.
The micro-sample test strip according to claim 1, wherein the sealing member is a mouth film, and the periphery of the sealing film is sealed and adhered to the periphery of the liquid discharge port.
A micro sample detecting test paper according to claim 5, wherein the groove cover is a grooved mask.
A micro sample detecting test paper according to claim 6, wherein said stable sample The fixed liquid storage tank is filled with blood stabilizing liquid.
A micro sample detecting test paper according to claim 7, wherein the blood stabilizing liquid is an EDTA buffer having a pH of 6.8 to 7.6.
A micro-sample test strip according to claim 1, wherein the anti-coagulation sample tube is a siphon tube containing an anticoagulant, one end of the siphon tube is connected to the negative pressure member, and the other end of the siphon tube is provided. There is a tube film.
A micro-sample test strip according to claim 9, wherein the anticoagulant is a 0.1-1.5 wt% heparin solution.