WO2021004091A1 - 一种毒物定性半定量检测试剂盒及毒物定性半定量方法 - Google Patents

一种毒物定性半定量检测试剂盒及毒物定性半定量方法 Download PDF

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WO2021004091A1
WO2021004091A1 PCT/CN2020/080795 CN2020080795W WO2021004091A1 WO 2021004091 A1 WO2021004091 A1 WO 2021004091A1 CN 2020080795 W CN2020080795 W CN 2020080795W WO 2021004091 A1 WO2021004091 A1 WO 2021004091A1
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qualitative
sample
semi
detection
tested
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French (fr)
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黄中伟
祁雷
孙华林
蒋海燕
顾志峰
梁桂文
奚庆华
肖明兵
吴征华
董岩松
毛国敏
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南通大学附属医院
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Priority to ZA2020/06353A priority Critical patent/ZA202006353B/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7756—Sensor type
    • G01N2021/7759—Dipstick; Test strip

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  • the invention relates to the field of research and development of medical rapid detection reagents and test papers, in particular to a toxicant qualitative and semi-quantitative detection kit and a toxicological qualitative method using the same, which is applied to clinical rapid toxicant detection.
  • the technical problem to be solved by the present invention is to provide a qualitative and semi-quantitative detection kit for poisons and a qualitative and semi-quantitative poison detection method.
  • the simple color reaction principle is used to prepare qualitative detection strips and semi-quantitative detection kits, which can eliminate paraquat poisoning in the clinic. Provide new diagnostic methods, effectively serve the clinic, and have extremely high social value.
  • the embodiments of the present invention provide a qualitative and semi-quantitative detection kit for poisons.
  • the kit is provided with a 5ml detection solution centrifuge tube containing 3ml detection solution, a 5ml blank centrifuge tube, and a The reaction color chart and a qualitative test reagent strip after adding the test solution at different concentrations of paraquat.
  • the present invention also provides a method for preparing a test solution.
  • a 5ml test tube add 3ml ddH 2 O, then add at least 0.1g of alkaline reagent and add at least 0.05g of sodium dithionite, wherein the alkaline reagent and The mass ratio of sodium disulfite is 2:1; shake well and store at room temperature.
  • the alkaline reagent is sodium bicarbonate.
  • the present invention also provides a qualitative and semi-quantitative method for poisons using the kit, which includes the following steps:
  • the sample to be tested is added to the test solution, and the mass ratio of the sample to be tested to the test solution is 1:4 to 4:1;
  • step (1) If the mixed solution in step (1) changes color, it means that the sample to be tested contains paraquat;
  • the sample to be tested is one of serum, plasma and urine.
  • the mass ratio of the sample to be tested and the test liquid is 1:1.
  • the present invention also provides a qualitative detection reagent strip for poisons, comprising a substrate on which there are a soaking end, a marking line, a detection line, and a hand-held end in sequence.
  • the area from the soaking end to the marking line and the detection line is provided with a
  • the sample cell, the capillary channel, and the reaction cell preset with alkaline reagent and sodium dithionite; the sample addition cell and the reaction cell are connected through the capillary channel, and the total number of alkaline reagents and sodium dithionite in the reaction cell is the same as that of the reaction cell Corresponding to the number.
  • the sample to be tested After the immersion end is immersed in the sample to be tested (urine, serum, plasma), the sample to be tested enters the sample addition tank and then enters the reaction tank through the capillary channel. Once the reaction tank changes color, it means that the test sample contains paraquat.
  • the beneficial effects of the above technical solutions of the present invention are as follows:
  • the present invention uses the principle of the color change reaction of sodium dithionite and paraquat under alkaline conditions to prepare qualitative test strips and semi-quantitative test kits, which can provide clinical treatment of paraquat poisoning
  • the new diagnosis method effectively serves the clinic and has extremely high social value.
  • Figure 1 is a color chart of the reaction after adding the test solution at different concentrations of paraquat
  • Figure 2 is a schematic diagram of the structure of the detection kit of the present invention.
  • Figure 3 is a schematic diagram of the structure of a qualitative detection reagent strip for poisonings of the present invention.
  • Figure 4 is a schematic diagram of the structure of the sample addition pool in the present invention.
  • an embodiment of the present invention provides a qualitative and semi-quantitative detection kit for poisons.
  • the kit is provided with a 5ml detection solution centrifuge tube containing 3ml detection solution, a 5ml blank centrifuge tube, and a The reaction color chart 10 and a qualitative test reagent strip after adding the test solution at different concentrations of paraquat.
  • the embodiment of the present invention also provides a method for preparing a test solution.
  • a 5ml test tube add 3ml ddH 2 O, then add more than 0.1g of alkaline reagent, and add more than 0.05g of sodium dithionite.
  • the alkaline reagent The mass ratio of and sodium dithionite is 2:1; shake well and can be stored for a long time at room temperature.
  • the alkaline reagent is sodium bicarbonate.
  • the embodiment of the present invention also provides a qualitative and semi-quantitative method for poisons using the above kit, which includes the following steps:
  • step (1) If the mixed solution in step (1) changes color, such as light yellow, light blue, blue or ink blue, it means that the sample to be tested contains paraquat;
  • Figure 1 shows the reaction color chart after dropping the test solution at different concentrations of paraquat.
  • the shade of gray and black in Figure 1 indicates the shade of blue after the test.
  • the detected concentration was above 1.5*10-5g/ml.
  • the reaction color chart we can also semi-quantitatively evaluate the concentration of paraquat in the sample to be tested. This situation has high clinical application value and significance for the treatment efficacy and prognosis evaluation of patients who are clinically diagnosed with paraquat poisoning.
  • Concentration assessment can comprehensively evaluate the efficacy and prognosis of the patient based on the color of the blood and urine sample's discoloration reaction, the time to gradually fade to colorless, and the patient's related symptoms, signs and auxiliary examination results.
  • the present invention also provides a qualitative detection reagent strip for poisons as shown in FIG. 3, comprising a substrate on which there are a soaking end 1, a marking line 2, a detection line 3, and a hand-held end 4 in sequence.
  • the soaking end 1 to The area of the marking line 2 and the detection line 3 is provided with a sample addition pool 5, a capillary channel 6, and a reaction pool 7 preset with alkaline reagents and sodium dithionite; the sample addition pool 5 and the reaction pool 7 pass through the capillary channel 6 Connected, as shown in Figure 4, the total number of alkaline reagents and sodium dithionite in the reaction tank 7 corresponds to the number of reaction tanks.
  • the sample to be tested After the immersion end is immersed in the sample to be tested (urine, serum, plasma), the sample to be tested enters the sample addition tank and then enters the reaction tank through the capillary channel. Once the reaction tank changes color, it means that the test sample contains paraquat.

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Abstract

一种毒物定性半定量检测试剂盒及毒物定性半定量方法，试剂盒内设有一个含有3ml检测液的5ml检测液离心管(8)、一个5ml的空白离心管(9)、一张不同百草枯浓度时加入检测液后的反应比色卡(10)及一根定性检测试剂条(11)。利用上述试剂盒的毒物定性半定量方法包括如下步骤：(1)待测样本加至检测液中，待测样本和检测液的质量比为1∶4～4∶1；(2)若步骤(1)中的混合液发生变色，代表待测样本中含有百草枯；(3)当利用定性检测试剂条(11)浸泡端(1)浸泡入待测样本中，反应池(7)出现变色代表待测样本中含有百草枯。可有效服务于临床排除百草枯中毒。

Description

一种毒物定性半定量检测试剂盒及毒物定性半定量方法 技术领域
本发明涉及医疗快速检测试剂、试纸研发领域，具体涉及一种毒物定性及半定量检测试剂盒及利用其的毒物定性方法，应用于临床快速毒物检测。
背景技术
目前全国针对毒物检测仅限于公安、农学中农药药物残留检测，在医学上仅有少数医院能够开展毒物检测，而且目前全国范围内医院中都没有针对毒物的快速毒物检测试剂盒，临床中毒判断效率低。
发明内容
本发明要解决的技术问题是提供一种毒物定性半定量检测试剂盒及毒物定性半定量方法，利用简单的变色反应原理，制备定性检测条、半定量检测试剂盒，能够为临床排除百草枯中毒提供新的诊断方法，有效地服务于临床，具有极高的社会价值。
为解决上述技术问题，本发明的实施例提供一种毒物定性半定量检测试剂盒，所述试剂盒内设有一个含有3ml检测液的5ml的检测液离心管、一个5ml的空白离心管、一张不同百草枯浓度时加入检测液后的反应比色卡及一根定性检测试剂条。
本发明还提供一种检测液的制备方法，于5ml试管中，加入3ml ddH 2O，再依次加入至少0.1g的碱性试剂，加入至少0.05g的连二亚硫酸钠，其中，碱性试剂和连二亚硫酸钠的质量比为2:1；摇匀，室温下保存。
优选的，所述碱性试剂为碳酸氢钠。
本发明还提供一种利用试剂盒的毒物定性半定量方法，包括如下步骤：
(1)待测样本加至检测液中，待测样本和检测液的质量比为1:4～4:1；
(2)若步骤(1)中的混合液发生变色，代表待测样本中含有百草枯；
(3)当利用定性检测条浸泡端浸泡入待测样本中，随即反应池出现变色，代表待测样本中含有百草枯。
其中，所述待测样本为血清、血浆和尿液中的一种。
优选的，所述待测样本和检测液的质量比为1:1。
本发明还提供一种毒物定性检测试剂条，包括一个基板，所述基板上依次为浸泡端、标记线、检测线、手持端，所述浸泡端到标记线及检测线的区域内设有加样池、毛细通道、预设有碱性试剂和连二亚硫酸钠的反应池；所述加样池、反应池通过毛细通道连通，所述反应池内的碱性试剂和连二亚硫酸钠的总数与反应池的数量相对应。当浸泡端浸泡到待测样本(尿液、血清、血浆)后，待测样本进入加样池，后通过毛细通道进入反应池，一旦反应池发生变色，代表待测样本中含有百草枯。
本发明的上述技术方案的有益效果如下：本发明利用连二亚硫酸钠和百草枯在碱性条件下发生变色反应的原理，制备定性检测条、半定量检测试剂盒，能够为临床排除百草枯中毒提供新的诊断方法，有效地服务于临床，具有极高的社会价值。
附图说明
图1为一张不同百草枯浓度时加入检测液后的反应比色卡；
图2为本发明中检测试剂盒的结构示意图；
图3为本发明中毒物定性检测试剂条的结构示意图；
图4为本发明中加样池处的结构示意图。
附图标记说明：
1、浸泡端；2、标记线；3、检测线；4、手持端；5、加样池；6、毛细通道；7、反应池；8、检测液离心管；9、空白离心管；10、反应比色卡；11、定性检测试剂条。
具体实施方式
为使本发明要解决的技术问题、技术方案和优点更加清楚，下面将结合具体实施例进行详细描述。
如图1所示，本发明实施例提供一种毒物定性半定量检测试剂盒，所述试剂盒内设有一个含有3ml检测液的5ml的检测液离心管、一个5ml的空白离心管、一张不同百草枯浓度时加入检测液后的反应比色卡10及一根定性检测试剂条。
本发明实施例还提供一种检测液的制备方法，于5ml试管中，加入3ml  ddH 2O，再依次加入0.1g以上的碱性试剂，加入0.05g以上的连二亚硫酸钠，其中，碱性试剂和连二亚硫酸钠的质量比为2:1；摇匀，室温下可长期保存。
优选的，所述碱性试剂为碳酸氢钠。
本发明实施例还提供一种利用上述的试剂盒的毒物定性半定量方法，包括如下步骤：
(1)取血清、血浆和尿液中的一种作为待测样本，将待测样本加至检测液中，待测样本和检测液的质量比为1:4～4:1，最佳质量比为1:1；
(2)若步骤(1)中的混合液发生变色，如变成淡黄色、淡蓝色、蓝色或者墨蓝色，代表待测样本中含有百草枯；
(3)当利用定性检测条浸泡端浸泡入待测样本中，随即反应池出现变色，代表待测样本中含有百草枯。
对于尿液本底颜色的影响：尿液本身呈黄色，深浅差别很大，为了消除这一影响，如果存在颜色过深，可以用1：1ddH 2O稀释后再行检测。
图1所示为不同百草枯浓度时，滴加本检测液后的反应比色卡，图1中灰黑度的深浅表示检测后蓝色的深浅。
实验过程中，检出浓度在1.5*10-5g/ml以上。根据该反应比色卡，我们还可以对待测样本进行百草枯浓度的半定量评估。这种情况，对于临床上确诊为百草枯中毒的患者治疗疗效及预后评估具有较高的临床应用价值及意义，我们可以对于百草枯中毒的患者，利用我们试剂盒，入院后进行每日的血尿浓度评估，根据血尿样本变色反应的颜色、逐步变淡直至无色的时间，以及患者相关症状、体征及辅助检查结果，可以综合评估患者疗效及预后情况。
本发明还提供一种如图3所示的毒物定性检测试剂条，包括一个基板，所述基板上依次为浸泡端1、标记线2、检测线3、手持端4，所述浸泡端1到标记线2及检测线3的区域内设有加样池5、毛细通道6、预设有碱性试剂和连二亚硫酸钠的反应池7；所述加样池5、反应池7通过毛细通道6连通，如图4所示，所述反应池7内的碱性试剂和连二亚硫酸钠的总数与反应池的数量相对应。当浸泡端浸泡到待测样本(尿液、血清、血浆)后，待测样本进入加样池，后通过毛细通道进入反应池，一旦反应池发生变色，代表待测样本中含有百草枯。
以上所述是本发明的优选实施方式，应当指出，对于本技术领域的普通技术 人员来说，在不脱离本发明所述原理的前提下，还可以作出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (7)

  1. 一种毒物定性半定量检测试剂盒，其特征在于，所述试剂盒内设有一个含有3ml检测液的5ml的检测液离心管、一个5ml的空白离心管、一张不同百草枯浓度时加入检测液后的反应比色卡及一根定性检测试剂条。
  2. 一种检测液的制备方法，其特征在于，于5ml试管中，加入3ml ddH 2O，再依次加入至少0.1g的碱性试剂，加入至少0.05g的连二亚硫酸钠，其中，碱性试剂和连二亚硫酸钠的质量比为2:1；摇匀，室温下保存。
  3. 根据权利要求2所述的检测液的制备方法，其特征在于，所述碱性试剂为碳酸氢钠。
  4. 一种利用权利要求1所述的试剂盒的毒物定性半定量方法，其特征在于，包括如下步骤：
    (1)待测样本加至检测液中，待测样本和检测液的质量比为1:4～4:1；
    (2)若步骤(1)中的混合液发生变色，代表待测样本中含有百草枯；
    (3)当利用定性检测条浸泡端浸泡入待测样本中，随即反应池出现变色，代表待测样本中含有百草枯。
  5. 根据权利要求4所述的毒物定性半定量方法，其特征在于，所述待测样本为血清、血浆和尿液中的一种。
  6. 根据权利要求4所述的毒物定性半定量方法，其特征在于，所述待测样本和检测液的质量比为1:1。
  7. 一种毒物定性检测试剂条，其特征在于，包括一个基板，所述基板上依次为浸泡端、标记线、检测线、手持端，所述浸泡端到标记线及检测线的区域内设有加样池、毛细通道、预设有碱性试剂和连二亚硫酸钠的反应池；所述加样池、反应池通过毛细通道连通，所述反应池内的碱性试剂和连二亚硫酸钠的总数与反应池的数量相对应。
PCT/CN2020/080795 2019-07-09 2020-03-24 一种毒物定性半定量检测试剂盒及毒物定性半定量方法 WO2021004091A1 (zh)

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