Multi-channel platelet aggregation capability whole blood detection reagent card

Abstract

The utility model belongs to the field of medical examination, and particularly relates to a multichannel platelet aggregation capability whole blood detection reagent card which comprises a sample inlet, a temporary storage bin, a microfluidic channel and a plurality of detection bins, the sample inlet is used for fixing a blood collection tube, and a sample is led to the temporary storage bin through the blood collection needle and the connecting tube; the bottom of the temporary storage bin is connected with the micro-fluidic channel and the sample flowing channel and further communicated with the N detection bins, and N is a positive integer and is greater than or equal to 2; the sample inlet is used for fixing the blood collection tube, the blood collection needle penetrates through the blood collection tube from the bottom, and a sample is guided to the temporary storage bin through a connecting tube at the tail of the blood collection needle; a sample flowing channel is arranged in each detection bin, and a sample in the temporary storage bin enters the detection bin through the micro-fluidic channel, the main liquid separation guide groove and the sample flowing channel in sequence. According to the utility model, clinical rapid detection can be met; the sample flow is accurately controlled; the detection process is simplified, only one tube of whole blood is extracted, and results of different activation pathways of the platelet aggregation function can be obtained without pretreatment.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the field of medical examination, especially relate to a kind of multi-channel platelet agglutination ability whole blood detection reagent card. BACKGROUND

[0002] The platelet agglutination detection reagent device currently applied on market mostly needs to use platelet plasma containing after centrifugation to detect.Because the density difference between platelet and red blood cell is small, plus the platelet content difference between different population (125~350×10 9 / L) is larger, it is difficult to achieve the standardization of platelet-poor plasma and platelet-rich plasma by centrifugation operation, and further affect the corresponding detection result.

[0003] The reagent card on the market includes sample bin, temporary storage bin and four detection bins, because the number of detection bin is small, to cope with different activation ways of platelet, it needs to extract blood sample multiple times or extract a large amount of blood sample at one time, for the patient of cardiovascular disease, the amount of blood extraction at one time is more important, and for operator, it increases the complexity of work, and because preparation and detection process time is longer, it cannot meet the instant requirement of clinic. UTILITY MODEL CONTENT

[0004] In view of the defects in prior art, the utility model provides a kind of multi-channel platelet agglutination ability whole blood detection reagent card, can meet the rapid detection of clinic;Blood sample flow is accurately controlled;Detection process is simplified, and only one tube of blood is extracted to obtain the result of different activation ways of platelet agglutination function.

[0005] The utility model is realized as follows:

[0006] A kind of multi-channel platelet agglutination ability whole blood detection reagent card, including sample inlet, temporary storage bin, microfluidic channel, main distribution channel, sample flow channel and N detection bin, N is positive integer, and N≥2;

[0007] The sample inlet is used to fix blood collection tube, blood collection needle is through blood collection tube by bottom, and sample is guided to temporary storage bin by connecting pipe of tail of blood collection needle;One sample flow channel is arranged in each detection bin, and sample in temporary storage bin enters detection bin by microfluidic channel, main distribution channel and sample flow channel in sequence.

[0008] Further, the relative two sides of temporary storage bin and each detection bin are equipped with light detection channel.

[0009] Further, stirring device for stirring sample is placed in each detection bin.

[0010] Further, the stirring device comprises a limiting baffle fixed horizontally on the inner wall of the detection bin and steel balls at the bottom of the limiting baffle; a gap exists between the limiting baffle and the inner wall of the detection bin so as to allow sample flow; the gap width is less than the diameter of the steel balls.

[0011] Further, the inside wall of the temporary storage bin is provided with blood absorption cotton at a specified height.

[0012] Further, detection cotton is arranged at the same position in each detection bin.

[0013] Further, the outer surface of the reagent card is provided with an information code label.

[0014] The utility model has the following technical effects:

[0015] The reagent card of the utility model comprises N (N≥2) detection bins, and different platelet activators are added in each detection bin, so that the activity detection of multiple platelet activation channels can be completed by extracting a small amount of blood at one time, the needs of the clinic are met, and a more comprehensive, more accurate, faster and more convenient guidance scheme is provided for the clinic diagnosis and treatment. The reagent card of the utility model can be matched with a platelet agglutination capacity detector to realize rapid, efficient and accurate detection. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 It is a principle diagram of the utility model platelet agglutination capacity detection platform;

[0017] Figure 2 It is a structural schematic diagram of the utility model four-channel reagent card;

[0018] Figure 3 It is an algorithm model diagram of the utility model detector;

[0019] Figure 4 It is a working principle schematic diagram of the utility model reagent card identification module;

[0020] Figure 5 It is a working principle schematic diagram of the utility model automatic sampling module;

[0021] Figure 6 It is a working principle schematic diagram of the utility model first infrared sensor receiving module;

[0022] Figure 7 It is a working principle schematic diagram of the utility model second infrared sensor receiving module;

[0023] Figure 8 It is a working principle schematic diagram of the utility model temperature control module, motor stirring module and light path detection module;

[0024] Figure 9It is the side view of the reagent card of the utility model.

[0025] Figure 10 It is the working principle schematic view of the calibration module.

[0026] In the figure: 1, reagent card;2, temporary storage warehouse;2a, blood absorption cotton;2b, first air hole;2c, second air hole;3, microfluidic channel;4, detection warehouse;4a, detection cotton;4b, limiting baffle;4c, steel ball;5, blood collection tube;6, information code label;7, sample inlet;8, light detection channel;9, sample flow channel;10, main liquid distribution guide groove;11, air groove;12, total exhaust hole;13, exhaust groove;14, exhaust small hole. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantage of the utility model more clearly, the following is combined with example, and the utility model is further detailedly explained.It should be understood that the specific embodiment described here is only used to explain the utility model, and is not used to limit the utility model.

[0028] As Figures 1-10 The utility model discloses a kind of multi-channel platelet agglutination capacity whole blood detection reagent cards, and the reagent card 1 includes sample inlet 7, temporary storage warehouse 2, microfluidic channel 3, main liquid distribution guide groove 10, sample flow channel 9 and N detection warehouses 4, N is positive integer, and N≥2.

[0029] The sample inlet 7 is used for fixing blood collection tube 5, and blood collection needle is through blood collection tube 5 by bottom, and sample is guided to temporary storage warehouse 2 by the connecting pipe of blood collection needle tail portion;Each detection warehouse 4 is equipped with a sample flow channel 9;Sample in temporary storage warehouse 2 sequentially passes through microfluidic channel 3, main liquid distribution guide groove 10 and sample flow channel 9 into detection warehouse 4;

[0030] The microfluidic channel 3 is equipped with first, second air hole 2c, and the first air hole 2b is connected with one-way valve;Second air hole 2c is connected with No. 1 electromagnetic valve and No. 2 electromagnetic valve through gas circuit, No. 1 electromagnetic valve and No. 2 electromagnetic valve are connected in parallel, and No. 1 electromagnetic valve is connected with the air inlet of vacuum pump, and No. 2 electromagnetic valve is connected with the air outlet of vacuum pump;

[0031] The designated height of the inner side wall of the temporary storage bin 2 is provided with blood absorption cotton 2a. When the blood absorption cotton 2a contacts the sample, discoloration occurs. The recognition module detects the change and sends a signal to the MCU slave processing module. Then, the control circuit controls the opening and closing of the vacuum pump, the first electromagnetic valve, the second electromagnetic valve and the one-way valve. When the temporary storage bin 2 needs a negative pressure environment to extract the sample in the blood collection tube 5, the one-way valve is closed, and no air flows into the first vent hole 2b. The second electromagnetic valve is closed, the first electromagnetic valve is turned on, and the vacuum pump controls the first electromagnetic valve to extract the air in the temporary storage bin 2 from the second vent hole 2c. When the sample in the temporary storage bin 2 reaches the height of the blood absorption cotton, the infrared sensor detected by the recognition module will send a detection signal to the MCU slave processing module. The first electromagnetic valve is closed, the second electromagnetic valve is turned on, and air enters the temporary storage bin 2 through the second vent hole 2c. As the sample is injected into each detection bin 4 through the microfluidic channel 3, air is discharged through the first vent hole 2b.

[0032] All the detection bins are provided with a common vent groove 11 at the top, and a total exhaust hole 12 is provided at the center of the vent groove 11 and communicates with the outside. Each detection bin is connected to the vent groove 11 through an exhaust groove 13, and an exhaust small hole 14 and detection cotton 4a opposite to the exhaust small hole are provided at the connection between the exhaust groove 13 and the vent groove 11. There is a gap between the detection cotton and the top and bottom of the exhaust groove for air flow;

[0033] The air in each detection bin is discharged to the outside through the exhaust small hole 14 and the total exhaust hole 12, respectively. When the sample in the detection bin is full and the air is discharged, the sample enters the exhaust groove until it contacts the detection cotton 4a. The detection cotton expands rapidly at the moment when the bottom of the detection cotton contacts the sample, closing the exhaust small hole. This detection bin cannot discharge gas and cannot continue to sample, but the top of the detection cotton has a gap with the exhaust groove, so it does not affect the gas flow of other detection bins. Therefore, even if the middle detection bin is full first, it does not affect the gas discharge of other detection bins. At the same time, the color of the expanded detection cotton changes; when the detection cotton of all detection bins expands and changes color, the infrared sensor detected by the recognition module will send a signal to the MCU slave processing module. Stop supplying power to the vacuum pump, the first electromagnetic valve and the second electromagnetic valve, and complete the sampling of all detection bins. Since the internal volume of each detection bin is the same and will be filled with sample, the flow rate of each detection bin is ensured to be the same.

[0034] In order to facilitate the uniform mixing and complete reaction of the sample and the platelet activator, a stirring device is placed in each detection bin 4. A magnet is provided on the detection motor drive shaft of the detector. When the motor rotates, it can drive the stirring device in the detection bin 4 to move in a certain track, thereby playing a stirring function for the reaction.

[0035] Preferably, the relative two sides of the temporary storage bin 2 and each detection bin 4 are provided with light detection channels 8 for light to pass through.

[0036] Preferably, the outer surface of the reagent card 1 is provided with an information code label 6 to facilitate scanning by the detector and acquisition of relevant information of the reagent card 1.

[0037] The following provides a detailed description of reagent card 1 through several examples:

[0038] Example 1

[0039] like Figure 2 As shown, this embodiment discloses a four-channel reagent card 1, which includes four independent detection compartments 4 arranged in sequence (I to IV) and connected to a sample storage compartment 2.

[0040] 1) Method 1: Using detection chamber III as the background channel:

[0041] Detection chamber III contains no detection reagents and serves as a blank control. The remaining three detection chambers I, II, and IV contain the same detection reagents as the corresponding detection chambers in "Method II" described below. The sample to be tested is introduced into the sample flow channel 9 of reagent card 1 through the injection port and flows along the sample flow channel 9 into the four detection chambers 4. The detection reagents in detection chambers I, II, and IV dissolve in the sample and then react with the detection components in the sample, producing a signal change relative to the blank control in sample detection chamber III. The magnitude of the signal change is used to determine the sample detection.

[0042] 2) Method 2: Using temporary storage bin 2 as the background channel:

[0043] Temporary storage chamber 2 does not contain test reagents and serves as a blank control. Each test chamber 4 contains a test result amplifier and a platelet activator. The platelet activator used in each test chamber is different. The test result amplifier is made of stained latex microspheres coated with fibrinogen. When the latex microspheres do not aggregate, the turbidity of test chamber 4 is high. Conversely, when they adhere to and aggregate with activated platelets, the turbidity of test chamber 4 decreases. The main function of the platelet activator is to activate platelets, thereby causing the remaining latex microspheres to aggregate and changing the turbidity of the sample in test chamber 4.

[0044] The detection chamber I contains a channel I detection reagent, the channel I detection reagent contains a lyophilized agent made of a detection result amplifying agent; the detection chamber II contains a channel II detection reagent, the channel II detection reagent contains a lyophilized agent made of a platelet activating agent and a detection result amplifying agent; the detection chamber III contains a channel III detection reagent, the channel III detection reagent contains a lyophilized agent made of a platelet activating agent and a detection result amplifying agent; the detection chamber IV contains a channel IV detection reagent, the channel IV detection reagent contains a lyophilized agent made of a platelet activating agent and a detection result amplifying agent. The sample to be detected enters the sample flow channel 9 of the reagent card 1 through the sample inlet, flows along the sample flow channel 9 to the four detection chambers 4, and the detection reagents in the detection chambers I, II, III and IV are dissolved in the sample, and then react with the detection components in the sample, thereby producing a signal change relative to the blank control in the sample temporary storage chamber 2, and the detection of the sample is completed by the size of the signal change.

[0045] Preferably, the platelet activating agent is selected from arachidonic acid, adenosine diphosphate, collagen, adrenaline, 5-hydroxytryptamine, prostaglandin, thromboxane A2, thrombin, thrombin-like enzyme, thrombin-like enzyme activating peptide, and any one of ristocetin, phospholipase A, phospholipase C, thrombin, thrombin-like enzyme, thrombin-like enzyme activating peptide. 2、 Preferably, the platelet activating agent is selected from arachidonic acid, adenosine diphosphate, collagen, adrenaline, 5-hydroxytryptamine, prostaglandin, thromboxane A2, thrombin, thrombin-like enzyme, thrombin-like enzyme activating peptide, and any one of ristocetin, phospholipase A, phospholipase C, thrombin, thrombin-like enzyme, thrombin-like enzyme activating peptide.

[0046] Introduction of the principle of platelet aggregation:

[0047] The role of adenosine diphosphate (ADP): When blood vessels are damaged, the damaged blood vessel wall and surrounding tissues release ADP. ADP can bind to receptors on the surface of platelets, triggering platelet activation and aggregation. The binding of ADP leads to the release of calcium ions within the platelet and the activation of intracellular signaling pathways, promoting changes in platelet morphology and function, making them more adhesive and aggregated.

[0048] The role of thrombin: During blood vessel injury, the damaged blood vessel wall releases tissue factor, activating the formation of thrombin. Thrombin can convert fibrinogen into fibrin, forming a blood clot. At the same time, thrombin can also directly stimulate platelet activation by binding to receptors on the surface of platelets, promoting platelet aggregation and morphological changes.

[0049] The role of phospholipase A and phospholipase C (PLA and PLC): Phospholipase is a class of enzymes that plays an important role in platelet activation. When platelets are activated, phospholipase is activated and translocated to the platelet surface, triggering phospholipid metabolism and changes. These changes can affect the properties of the cell membrane, making platelets more adhesive and aggregated.

[0050] The role of arachidonic acid: Arachidonic acid can be metabolized by enzymes in platelets into a series of bioactive substances, such as prostaglandins, thromboxane, etc. These substances can promote the activation and aggregation of platelets by activating intracellular signaling pathways, thereby participating in the process of thrombosis.

[0051] The role of collagen (COL): Collagen is an important extracellular matrix protein of vascular endothelial cells, which is exposed to blood and binds to receptors on the surface of platelets, thereby triggering the activation and aggregation of platelets. Collagen binds to receptors GPVI and α2β1 on the surface of platelets, activates platelets and induces them to release platelet activation factors such as platelet kinase and 5-hydroxytryptamine, thereby promoting the aggregation and morphological changes of platelets. In addition, collagen can further promote the activation and aggregation of platelets by activating the signal transduction pathways of platelets, such as phospholipase C and protein kinase C, etc.

[0052] The role of thromboxane A2: It is a bioactive substance produced by the metabolism of arachidonic acid. Thromboxane A2 mainly promotes the activation and aggregation of platelets by binding to the receptor TP on the surface of platelets. Thromboxane A2 activates intracellular signaling pathways in platelets, such as increasing intracellular calcium ion concentration, activating phospholipase C, etc., leading to changes in platelet morphology and release of platelet activation factors, thereby promoting platelet aggregation and thrombosis.

[0053] In summary, adenosine diphosphate, thrombin, phospholipase, arachidonic acid, collagen, thromboxane A2 can all activate platelets through different pathways and cause changes in their morphology and function, thereby participating in the process of thrombosis and hemostasis. The interaction and regulation of these biochemical reactions are important mechanisms of platelet activation and thrombosis.

[0054] The blood collection tube 5 contains an anticoagulated sample, which is undiluted whole blood, plasma or diluted whole blood, mixed with a sodium citrate solution at a volume ratio of 1:9, and the concentration of the sodium citrate solution is 0.109 mol / L or 0.129 mol / L.

[0055] Preferably, the detection chamber I contains a detection result amplifying agent, and the sample does not adhere to and aggregate with the latex microspheres in the detection chamber, so that the turbidity of the sample in the detection chamber I is the largest;

[0056] Preferably, the detection bin II is activated by one of the platelet activators, i.e. the thrombin-like enzyme, which activates the platelets directly to cause a cascade reaction, so that the sample and the latex microspheres in the detection bin adhere and aggregate, thereby changing the turbidity of the sample in the detection bin II, and the maximum aggregation of the platelets under the thrombin-like enzyme is obtained, and the corresponding results are obtained through the cooperation of the specific hardware and software in the device.

[0057] Preferably, the detection bin III is used as a background channel in the following two ways:

[0058] The first way is to use the detection bin III as a background channel: the detection bin III does not contain a detection reagent, and is used as a blank control.

[0059] The second way is to use the temporary storage bin 2 as a background channel: the detection bin III is one of the platelet activators, i.e. the mixture of phospholipase A and phospholipase C or arachidonic acid; the mixture of phospholipase A and phospholipase C or arachidonic acid activates the platelets and releases more platelet aggregation factors, thereby causing a cascade reaction, so that the sample and the latex microspheres in the detection bin adhere and aggregate, thereby changing the turbidity of the sample in the detection bin III. If the subject has taken a drug such as a cyclooxygenase-1 inhibitor (e.g. low-dose 81-325mg aspirin, indobufen, etc.) before, the activation of the platelets will be inhibited, the turbidity change rate in the detection bin will be reduced, and the corresponding results obtained through the cooperation of the specific hardware and software in the device will also be reduced.

[0060] Preferably, the detection bin IV is one of the platelet activators, i.e. adenosine diphosphate. Adenosine diphosphate can bind to the adenosine diphosphate receptor on the surface of the platelet membrane, activate the platelets, and release thrombin and adenosine diphosphate and other substances, thereby causing a cascade reaction, so that the sample and the latex microspheres in the detection bin adhere and aggregate, thereby changing the turbidity of the sample in the detection bin IV. If the subject has taken an ADP inhibitor (e.g. clopidogrel) before, the activation of the platelets will be inhibited, the turbidity change rate in the detection bin will be reduced, and the corresponding results obtained through the cooperation of the specific hardware and software in the device will also be reduced.

[0061] The reagent card 1 can realize the detection of the platelet aggregation ability, and the detection method is as follows:

[0062] 1) Calculate the platelet aggregation rate AggI of the detection bin I by using the matching software;

[0063] 2) Calculate the platelet aggregation rate AggII of the detection bin II by using the matching software;

[0064] 3) Calculate the platelet aggregation rate AggIII of the detection bin III by using the matching software;

[0065] 4) Calculate the platelet aggregation rate (AggⅣ) in the detection chamber using the accompanying software.

[0066] The software algorithm is as follows:

[0067] like Figure 3 As shown, the software algorithm uses a mathematical model fitting method. Based on the differences in the detection reagents set in each detection chamber, the platelet aggregation reaction process is first established as a curve with time t as the independent variable and light transmittance as the dependent variable. The algorithm formula is then fitted based on Lambert-Beer's law and the change in microsphere concentration.

[0068] Figure 3 The meaning of the curves in the image is as follows:

[0069] Blood whole blood sample reaction line: The testing chamber or temporary storage chamber it contains contains no latex microspheres or activators. Platelets are not activated and there are no latex microspheres or activators to obstruct light transmission through the sample, so the light transmittance is the highest;

[0070] Blood+Mic, or whole blood sample + latex microsphere reaction line: After the sample enters the detection chamber, it is stirred and mixed with the latex microspheres. Since there is no activator, platelets are not activated and will not adhere to or aggregate with the latex microspheres. At the same time, the latex microspheres have a dye on their surface, which blocks the detection light. Therefore, this detection chamber has the lowest light transmittance and can be used as a low-value blank control.

[0071] TRAP, ADP, PLA+PLA, COL, and COX-1 are listed in parallel; they are all different types of activators used in the detection chamber. Each activates platelets through its respective pathway, causing them to adhere and aggregate with latex microspheres in the detection chamber, thereby allowing for the calculation of light transmittance.

[0072] in:

[0073] TRAP: Thrombin receptor activating peptide activation response curve;

[0074] ADP: Adenosine diphosphate activator activation reaction curve;

[0075] PLA+PLC: Activation reaction curves of phospholipase A and phospholipase C;

[0076] AA: Arachidonic acid activation reaction curve;

[0077] COL: Collagen activation reaction curve;

[0078] COL-1: Cyclooxygenase-1 inhibitor curve;

[0079] The temporary storage or detection chamber containing the blood corresponds to a maximum light transmittance of I. BloodSo its concentration is the lowest;

[0080] The light transmittance of the detection chamber where TRAP, ADP, PLA+PLA, COL and COX-1 are located is I TRAP , I ADP , I 2P , I COL and I AA ;

[0081] The minimum light transmittance I Blood+Mic corresponds to the detection chamber where Blood+Mic is located, so its concentration is the highest.

[0082] The concentration of the ADP curve is denoted as C ADP , C ADP =a*lg(I Blood / I ADP );

[0083] The maximum concentration is denoted as C ADP.MAX , C ADP.MAX =a*lg(I Blood / I ADP.MIN );

[0084] The concentration of the TRAP curve is denoted as C TRAP , C TRAP =a*lg(I Blood / I TRAP );

[0085] The maximum concentration is denoted as C TRAP.MAX , C TRAP.MAX =a*lg(I Blood / I TRAP.MIN );

[0086] The concentration of the PLA+PLC curve is denoted as C 2P , C 2P =a*lg(I Blood / I 2P );

[0087] The maximum concentration is denoted as C 2P.MAX , C 2P.MAX =a*lg(I Blood / I 2P.MIN );

[0088] The concentration of the COL curve is denoted as C COL , C COL =a*lg(I Blood / I COL );

[0089] The maximum concentration is denoted as C COL.MAX , C COL.MAX =a*lg(IBlood / I COL.MIN );

[0090] The concentration of COX-1 in the curve is denoted as C. AA C AA =a*lg(I Blood / I AA );

[0091] Its maximum concentration is denoted as C. AA.MAX C AA.MAX =a*lg(I Blood / I AA.MIN );

[0092] The concentration of blood is denoted as C. Blood =a*lg(I Blood / I Blood ) = 0;

[0093] The concentration of Blood+Mic is denoted as C. Blood+Mic =a*lg(I Blood / I Blood+Mic );

[0094] The temporary storage or detection chamber containing the blood corresponds to a maximum light transmittance of I. Blood Its concentration is the lowest;

[0095] The transmittance of the detection chambers containing TRAP, ADP, PLA+PLA, COL, and COX-1 are respectively I TRAP I ADP I 2P I COL and I AA The minimum transmittance of each of their detection chambers is I. TRAP.MIN I ADP.MIN I 2P.MIN I COL.MIN and I AA.MIN ;

[0096] The minimum light transmittance I corresponds to the detection chamber containing Blood+Mic. Blood+Mic Its concentration is the highest;

[0097] In summary, based on the configuration of the test reagents in the reagent card's testing compartment:

[0098] Platelet aggregation response unit PU in the P2Y12 receptor activation pathway P2Y12 :

[0099] PU P2Y12 =a*[(C MAX -C ADP ) / (C MAX -C Blood )]+b;

[0100] C MAX =C Blood+Mic or C ADP.MAX ;

[0101] Platelet aggregation response unit PU of the thrombin receptor activation pathway BASE :

[0102] PU BASE =a*[(C MAX -C TRAP ) / (C MAX -C Blood )]+b;

[0103] C MAX =C Blood+Mic or C TRAP.MAX ;

[0104] Platelet aggregation response unit AU of the cyclooxygenase-1 activation pathway AA :

[0105] AU AA =a*[(C MAX -C AA ) / (C MAX -C Blood )]+b;

[0106] C MAX =C Blood+Mic or C AA.MAX ;

[0107] Platelet aggregation response unit CU of the collagen activation pathway COL :

[0108] CU COL =a*[(C MAX -C COL ) / (C MAX -C Blood )]+b;

[0109] C MAX =C Blood+Mic or C COL.MAX ;

[0110] Platelet aggregation response unit PU of the phospholipase A and phospholipase C activation pathway 2P :

[0111] PU 2P =a*[(C MAX -C 2P ) / (C MAX -C Blood )]+b;

[0112] C MAX = C Blood+Mic or C 2P.MAX ;

[0113] In the above formula, a and b are constant terms; C MAX is a variable, which represents the maximum concentration reference parameter used in calculating the platelet aggregation reaction unit, as explained below:

[0114] 1) One of the detection chambers of the reagent card is configured only with a detection result amplifying agent, and its corresponding test concentration is C Blood+Mic ; when calculating the platelet aggregation unit of other detection chambers, this is taken as the maximum concentration reference parameter C MAX ;

[0115] 2) The reagent card is configured with both a platelet activator and a detection result amplifying agent in all detection chambers; when calculating the platelet aggregation unit of the detection chambers, the maximum test concentration of each detection chamber is used as the reference parameter C MAX .

[0116] Example 2

[0117] This embodiment discloses a five-channel reagent card 1, which comprises five detection chambers 4 arranged in sequence, each of which is independent, and a sample temporary storage chamber 2.

[0118] 1) Method one uses detection chamber III as a background channel

[0119] Detection chamber III does not contain a detection reagent, serving as a blank control; the remaining four detection chambers I, II, IV and V contain detection reagents, which are exactly the same as the corresponding detection chambers in the following "Method two". The sample to be tested enters the sample flow channel 9 of the reagent card 1 through the sample inlet, flows along the sample flow channel 9 to the five detection chambers 4, and the detection reagents in the detection chambers I, II, IV and V dissolve in the sample, immediately react with the detection components in the sample, produce signal changes relative to the blank control in the sample detection chamber III, and complete the detection of the sample through the signal change size.

[0120] 2) Method two uses temporary storage chamber 2 as a background channel

[0121] The temporary storage chamber 2 does not contain a detection reagent, serving as a blank control; each detection chamber 4 contains a detection result amplifying agent and a platelet activator; the platelet activators used in each detection chamber are different; the detection result amplifying agent is selected as dyed latex microspheres, the surface of which is coated with fibrinogen.

[0122] The detection chamber I contains channel I detection reagent, the channel I detection reagent contains lyophilized agent made of detection result amplifying agent; the detection chamber II contains channel II detection reagent, the channel II detection reagent contains lyophilized agent made of blood platelet activating agent and detection result amplifying agent; the detection chamber III contains channel III detection reagent, the channel III detection reagent contains lyophilized agent made of blood platelet activating agent and detection result amplifying agent; the detection chamber IV contains channel IV detection reagent, the channel IV detection reagent contains lyophilized agent made of blood platelet activating agent and detection result amplifying agent; the detection chamber V contains channel V detection reagent, the channel V detection reagent contains lyophilized agent made of blood platelet activating agent and detection result amplifying agent. The sample to be detected is introduced into the sample flow channel 9 of the reagent card 1 through the sample inlet, and the sample flows along the sample flow channel 9 to the five detection chambers 4, the detection reagents in the detection chambers I, II, III, IV and V are dissolved in the sample, the detection reagents react with the detection components in the sample, and a signal change is generated relative to the blank control in the sample temporary storage chamber 2, and the detection of the sample is completed through the signal change.

[0123] Preferably, the blood platelet activating agent is selected from any one of arachidonic acid, adenosine diphosphate, collagen, adrenaline, 5-hydroxytryptamine, prostaglandin, thromboxane A2, ristocetin, phospholipase A, phospholipase C, thrombin, isothrombin, isothrombin activating peptide;

[0124] Preferably, the detection chamber I is the detection result amplifying agent;

[0125] Preferably, the detection chamber II is one of the blood platelet activating agents, i.e. isothrombin activating peptide;

[0126] Preferably, the detection chamber III is selected from the following two ways:

[0127] The first way uses the detection chamber III as a background channel: the detection chamber III does not contain detection reagent, and is used as a blank control;

[0128] The second way uses the temporary storage chamber 2 as a background channel: the detection chamber III contains a mixture of phospholipase A and phospholipase C or arachidonic acid, which is one of the blood platelet activating agents;

[0129] Preferably, the detection chamber IV contains adenosine diphosphate, which is one of the blood platelet activating agents;

[0130] Preferably, the detection chamber V contains ristocetin, which is one of the blood platelet activating agents.

[0131] Finally, the device gives a quantitative value according to the change of the whole sample turbidity of different channels:

[0132] The channel I blood platelet aggregation rate AggI is calculated by using the matching software;

[0133] The software calculates the platelet aggregation rate Agg II of channel II;

[0134] The software calculates the platelet aggregation rate Agg III of channel III;

[0135] The software calculates the platelet aggregation rate Agg IV of channel IV;

[0136] The software calculates the platelet aggregation rate Agg V of channel V;

[0137] The software algorithm is the same as that of Example 1, which is not repeated here.

[0138] Example 3

[0139] Based on Example 1 and Example 2, a six-channel, seven-channel or more channel reagent card is prepared according to the needs of the clinic. The reagent card 1 includes six or seven or more detection chambers 4 respectively. The reagents of the first five channels of the six-channel reagent card 1 are consistent with those of the five-channel reagent card 1, that is, based on the detection items of the five-channel card, by adding a different platelet activator in the sixth detection chamber 4 than in other detection chambers 4, more detection requirements are realized.

[0140] The seven-channel reagent card 1 works similarly to the above description, the first six detection chambers 4 are consistent with the six-channel reagent card 1, and by adding a different platelet activator in the seventh detection chamber 4 than in other detection chambers, the detection requirements are further expanded.

[0141] The more-channel reagent card 1 is similar, which is not repeated here.

[0142] In summary, a small amount of blood is extracted at one time to complete the results required by the clinic, and more accurate, faster and more convenient guidance schemes are provided for the diagnosis and treatment of the clinic.

[0143] In order to further understand the working principle of the reagent card, the detailed structure of the platelet ability detector is introduced as follows:

[0144] The methodology basis of the platelet ability detector is the optical turbidimetry, also known as the turbidity determination method, which is a method for measuring the light intensity through the suspended particle medium to determine the concentration of suspended matter, and is a world-recognized method for detecting platelet function. The optical turbidimetry utilizes the change of the concentration of related light-absorbing substances in a solution to affect the absorbance of a solution to light of a specific wavelength, that is, the higher the concentration of light-absorbing substances, the higher the absorbance of the solution to light of a specific wavelength, and the lower the light transmittance, and vice versa, the lower the concentration of light-absorbing substances, the lower the absorbance of the solution to light of a specific wavelength, and the higher the light transmittance, so that the concentration of the specific light-absorbing substance can be determined and related analysis can be carried out. The stability of the light source of the platelet ability detector and the measurement accuracy of the light flux output thereof ultimately affect the accuracy of the results. Only by ensuring the stability of the light source driving and the accuracy of the light signal acquisition can the risk be minimized. In view of the above risks, the platelet ability detector is innovated as follows:

[0145] The detector comprises a reagent card containing bin, a control module, an automatic sampling module, a motor stirring module, a temperature control module, an optical path detection module, a calibration module, a data display and printing module;

[0146] Specifically,

[0147] The control module comprises an MCU slave processing module and a control circuit connected electrically;

[0148] The automatic sampling module comprises a reagent card recognition module, a vacuum pump, a first electromagnetic valve, a second electromagnetic valve, a one-way valve, a first infrared sensor receiving module, a second infrared sensor receiving module and a pressure sensor;

[0149] As shown in Figure 4 The reagent card recognition module comprises an information code label 6 attached to the reagent card 1 and a code scanner provided on the detector; the code scanner is used for scanning the information code label 6 on the reagent card 1 and outputting information to the control circuit, and the control circuit further transmits data to the MCU slave processing module.

[0150] As shown in Figure 5As shown, the detection bin 4 is connected with a one-way valve through the first vent hole 2b of the microfluidic channel; the temporary storage bin 2 is connected with a first electromagnetic valve and a second electromagnetic valve through the second vent hole 2c of the microfluidic channel and the gas path respectively, the exhaust port of the vacuum pump is connected with the second electromagnetic valve for the inflation of the temporary storage bin 2; the air inlet of the vacuum pump is connected with the first electromagnetic valve for the exhaust of the temporary storage bin 2; the control circuit can control the operation of the vacuum pump; the pressure sensor is used to detect the air pressure on the gas path of the first electromagnetic valve to determine whether there is an abnormality in the vacuumizing process; when the negative pressure environment is needed in the temporary storage bin 2 to extract the sample in the blood collection tube 5, the one-way valve is closed and no air flows into the first vent hole 2b; the second electromagnetic valve is closed and the first electromagnetic valve is turned on, and the vacuum pump controls the first electromagnetic valve to exhaust the air in the temporary storage bin 2 from the second vent hole 2c; when the sample in the temporary storage bin 2 reaches the set height, the first electromagnetic valve is closed and the second electromagnetic valve is turned on, and the air enters the temporary storage bin 2 through the second vent hole 2c; the one-way valve is opened, and as the sample is injected into each detection bin 4 through the microfluidic channel 3, the air is exhausted through the first vent hole 2b.

[0151] As shown in the figure, the first infrared sensor receiving module detects the color change of the blood absorption cotton 2a on the temporary storage bin 2 through an infrared sensor and feeds back a signal to the MCU slave processing module, and the MCU slave processing module sends an instruction to the control circuit to determine the working state of the vacuum pump according to the signal. Figure 6 As shown in the figure, the MCU slave processing module sends an inflation instruction to the control circuit, and the vacuum pump inflates the temporary storage bin 2 through the second electromagnetic valve, and the temporary storage bin 2 changes from negative pressure to positive pressure; at this time, since each detection bin channel of the reagent card 1 is under negative pressure, the sample liquid (whole blood or quality control liquid) flows into each detection bin 4. The detection cotton 4a is arranged at the same position in each detection bin 4; when the sample height reaches the detection cotton 4a, the color of the detection cotton 4a changes; the second infrared sensor receiving module monitors the color change of the detection cotton 4a through a plurality of infrared sensors corresponding to the detection bin 4 one by one and outputs a signal to the MCU slave processing module, and sends a closing inflation instruction to the control circuit to stop continuing to inject the sample into the detection bin 4.

[0152] Figure 7 According to the above structure, the working steps of the automatic sampling module are briefly described as follows:

[0153] Step 1) Reagent card 1 judgment:

[0154] Step 1) Reagent card 1 judgment:

[0155] ​After the information code label 6 is pasted to the prescribed area of the reagent card 1, the reagent card 1 is placed in the reagent card accommodating bin of the platform; after the test is started, the MCU slave processing module controls the power-on operation of the code scanner of the device, reads the label information of the reagent card 1, and outputs the information to the MCU slave processing module; after the MCU slave processing module receives the information of the reagent card 1, performs data analysis and judgment, and executes the corresponding detection program.

[0156] Step 2) sample injection of the temporary storage bin 2 of the reagent card 1:

[0157] The MCU slave processing module outputs the instruction of vacuum pumping, detects whether the pressure of the gas circuit is normal through the pressure sensor, judges whether the gas circuit is abnormal, and controls the circuit to make the vacuum pump power on and start vacuum pumping of the temporary storage bin 2, and the first electromagnetic valve power on and change from the closed state to the open state. The pressure sensor converts the detected air pressure into an electric signal and outputs the signal to the MCU slave processing module through the control circuit. After vacuum pumping, the negative pressure in the temporary storage bin 2 of the reagent card 1 is much lower than the air pressure in the blood collection tube 5 inserted into the reagent card 1, and the sample (whole blood or quality control liquid) in the blood collection tube 5 starts to flow to the temporary storage bin 2 of the reagent card 1.

[0158] Step 3) stopping of sample injection of the temporary storage bin 2 of the reagent card 1

[0159] When the liquid injection amount reaches the specified position of the temporary storage bin 2 of the reagent card 1, the blood absorption cotton 2a changes color. The first infrared sensor receiving module feeds back the signal of the color change of the blood absorption cotton 2a to the MCU slave processing module through the corresponding infrared sensor, and the MCU slave processing module sends an instruction to the control circuit according to the signal to stop the power supply of the vacuum pump and the electromagnetic valve 1, and completes the sample injection of the temporary storage bin 2.

[0160] Step 4) sample injection of the detection bin 4 of the reagent card 1:

[0161] The MCU slave processing module outputs the instruction of air pumping, detects whether the pressure of the gas circuit is normal through the pressure sensor, judges whether the gas circuit is abnormal, and controls the circuit to make the vacuum pump power on and start air pumping, and the first electromagnetic valve power off and close, and the second electromagnetic valve power on and open. The vacuum pump pumps air into the temporary storage bin 2 through the second electromagnetic valve, and the pressure in the temporary storage bin 2 changes from negative pressure to positive pressure. At this time, since each detection bin channel of the reagent card 1 is negative pressure, the sample liquid (whole blood or quality control liquid) flows to each detection bin 4.

[0162] Step 5) stopping of sample injection of the detection bin 4 of the reagent card 1:

[0163] After all the detection chambers 4 are filled with samples, the corresponding detection cotton 4a inside each detection chamber 4 will change color; the output signal of the corresponding infrared sensor will also change, and the MCU slave processing module will be notified; after receiving the infrared sensor signal, the MCU slave processing module will send a command to the control circuit to stop the power supply of the vacuum pump and the electromagnetic valve, and the sample loading of the detection chamber 4 will be completed.

[0164] As shown in Figure 8 The temperature control module includes a heating resistor and a thermal detection element. The reagent card containing chamber contains a heat-conducting aluminum block, which is attached to one side of the reagent card 1 and covers the temporary storage chamber 2 and each detection chamber 4. The heating resistor and the thermal detection element are fixed on the heat-conducting aluminum block, and heat is conducted through the aluminum block to control the temperature of the sample in the temporary storage chamber 2 and each detection chamber 4. After the instrument is turned on, the MCU slave processing module of the control module outputs the control signal of the heating circuit at the preset PWM frequency, supplies power to the heating resistor, and feeds back the received signal of the thermal detection element to the MCU slave processing module for temperature closed-loop feedback control processing, so that the detection sample of the reagent card 1 is maintained at a specified temperature.

[0165] To more accurately reproduce the "gold standard" light turbidimetry, the following technical aspects of basic work and improvements are made in cooperation with the reagent card 1 equipment. First, the accurate emission of photons in the device is realized by using precise optical devices and accurate control systems, including the use of specific LED light sources to ensure that the light source in the device has stable output power and wavelength, so that the emission of photons is stable and controllable. Second, the device captures photons. The photodiode detector used by the device has high sensitivity and low noise to ensure that the capture of photons is efficient and reliable. In addition, precise optical lenses and filter devices are also used to ensure the controllability of the capture direction and wavelength range of the photons.

[0166] As shown in Figure 8As shown, the optical path detection module includes a light source emitting module and a light source receiving module. Reagent card 1 is fixed in the reagent card receiving compartment and corresponds to the light source emitting module and the light source receiving module, with reagent card 1 perpendicular to the detection light beam. The MCU slave processing module powers on each of the light-emitting LEDs (corresponding one-to-one with the light detection channels 8) of the light source emitting module. The light-emitting LEDs emit light signals to each detection compartment 4. Each of the photodiodes (corresponding one-to-one with the light detection channels 8) of the light receiving module filters, amplifies, and converts the light signals transmitted through each light detection channel 8 into photoelectric signals, which are then transmitted to the MCU slave processing module. The processing module generates detection results such as AU or PU, which are displayed on the device screen. Specifically, the MCU slave processing module of the control module outputs a pre-sampling command, gradually increasing the power supply voltage of the predetermined channel circuit of the light source board until the corresponding channel receiving value of the light source receiving module reaches the preset standard. Then, based on the stored calibration compensation data, it supplies power to the remaining light source emitting circuits of the light source emitting module to begin the optical path detection operation. The lower-level software of this invention ensures the stable driving of the light source through a 12-bit DAC current source, and at the same time, it acquires and analyzes the spectral signal through a 12-bit ADC module, a photodiode, and a 3.3V reference source, thus ensuring the accurate acquisition of the optical signal and minimizing the risk.

[0167] like Figure 9 As shown, the motor stirring module includes a stirring device placed in each detection chamber 4 and a detection motor located below the reagent card 1. The stirring device includes a limiting baffle 4b horizontally fixed to the inner wall of the detection chamber and a steel ball 4c at its bottom. A magnet is provided on the drive shaft of the detection motor, so that when the motor rotates, it can drive the steel ball in the detection chamber 4 to move in a certain trajectory to stir the sample evenly. There is a gap between the limiting baffle 4b and the inner wall of the detection chamber to allow sample flow. The width of this gap is smaller than the diameter of the steel ball 4c, so that the steel ball is restricted to the bottom of the detection chamber and can only move horizontally. At the same time, the speed data of the stirring motor is fed back to the MCU slave processing module through the sensor on the corresponding detection motor drive shaft handle on the light source board for closed-loop feedback control of the stirring frequency.

[0168] like Figure 10As shown, the calibration module comprises an optical attenuation sheet, an electronic quality control card, a data storage module and a PC host computer; the optical attenuation sheet is fixed to the electronic quality control card and is fixed in the middle of the light source emitting module and the light source receiving module as a whole through the reagent card containing bin, and the optical attenuation sheet is vertically corresponding to each light emitting LED and photosensitive receiving diode respectively; the working principle of the module is as follows: the PC host computer software sends a calibration instruction, the MCU slave processing module starts the calibration processing software, first gradually increases the power supply voltage of the light source board pre-sampling channel circuit, so that the corresponding channel receiving value of the light source receiving module reaches the preset standard; then adjust the power supply of the other several light sources respectively, so that the corresponding receiving channels also reach the preset standard; finally, the corresponding relationship between the power supply of the other several light sources of the light source board and the pre-sampling channel power supply at this time is calculated respectively by the prior art, the calibration compensation data is obtained and transmitted to the data storage module.

[0169] The full-automatic platelet aggregometer applies the principle of optical turbidimetry, and when detecting, the working principle of the automatic sampling module is as follows: the information code on the reagent card 1 is scanned and recognized by the reagent card recognition module; then the automatic extraction and sampling of the blood sample are realized according to the obtained information, and the combination with the gas circuit, vacuum pump and corresponding electromagnetic valve of the platform and the liquid circuit structure of the reagent card 1 is ensured, so that the samples entering the detection bin are all unactivated samples; the motor stirring module is controlled by the control module to output the instruction of the detection motor according to the preset stirring frequency of the software, and the rotation of the detection motor stirs the reaction in the detection bin 4; the temperature control module is used to simulate the in-vivo reaction environment to ensure the stability of the surrounding temperature during in-vitro detection, and the corresponding polymerization reaction occurs in the different reaction bins of the reagent card 1 during the reaction process; the light path detection module detects the light value change caused by the polymerization reaction and calculates to obtain the experimental result; the calibration module ensures that the power supply voltage of the light path detection module can reach the preset standard through calibration compensation; and the data display and printing module realizes the display and printing functions of the final experimental result.

[0170] The above only describes the preferred embodiments of the present application, and is not used to limit the present application, and any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A multi-channel platelet aggregometry whole blood test kit, characterized by: The device comprises a sample inlet, a temporary storage bin, a microfluidic channel, a main distribution guide, a sample flow channel and N detection bins, wherein N is a positive integer and N≥2. The sample inlet is used for fixing a blood collection tube, a blood collection needle penetrates the blood collection tube from the bottom, and a connecting tube at the tail of the blood collection needle guides the sample to the temporary storage bin; each detection bin is provided with a sample flow channel, and the sample in the temporary storage bin enters the detection bin through the microfluidic channel, the main distribution guide and the sample flow channel in sequence.

2. The multi-channel platelet aggregability whole blood test kit according to claim 1, wherein: The temporary storage bin and each detection bin are provided with light detection channels on opposite sides.

3. The multi-channel platelet aggregability whole blood test kit as claimed in claim 1, wherein: Each detection bin is provided with a stirring device for stirring the sample.

4. The multi-channel platelet aggregability whole blood test kit as claimed in claim 3, wherein: The stirring device comprises a limiting baffle fixed horizontally on the inner wall of the detection bin and steel balls at the bottom of the limiting baffle; a gap exists between the limiting baffle and the inner wall of the detection bin for the flow of the sample; and the width of the gap is smaller than the diameter of the steel balls.

5. The multi-channel platelet aggregability whole blood test kit as claimed in claim 1, wherein: A blood absorption cotton is arranged on the inner side wall of the temporary storage bin at a specified height.

6. The multi-channel platelet aggregability whole blood test kit as claimed in claim 1, wherein: Detection cottons are arranged at the same position in each detection bin.

7. The multi-channel platelet aggregability whole blood test kit as claimed in claim 1, wherein: An information code label is arranged on the outer surface of the reagent card.

Images