WO2014180172A1 - 生物芯片及其应用 - Google Patents

生物芯片及其应用 Download PDF

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Publication number
WO2014180172A1
WO2014180172A1 PCT/CN2014/070289 CN2014070289W WO2014180172A1 WO 2014180172 A1 WO2014180172 A1 WO 2014180172A1 CN 2014070289 W CN2014070289 W CN 2014070289W WO 2014180172 A1 WO2014180172 A1 WO 2014180172A1
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WIPO (PCT)
Prior art keywords
substrate
reaction
biochip
film
detecting unit
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PCT/CN2014/070289
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English (en)
French (fr)
Inventor
董建国
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上海派莱星生物技术有限公司
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Application filed by 上海派莱星生物技术有限公司 filed Critical 上海派莱星生物技术有限公司
Priority to CN201490000637.4U priority Critical patent/CN205449989U/zh
Publication of WO2014180172A1 publication Critical patent/WO2014180172A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
    • A61B10/0045—Devices for taking samples of body liquids
    • A61B2010/0074—Vaginal or cervical secretions

Definitions

  • the invention relates to the technical field of clinical diagnosis medical instruments, in particular to a biochip and an application thereof.
  • vaginal secretions usually require the first collection of vaginal secretions, oral mucus, gastrointestinal secretions. Wait for the clinical liquid sample, and then take the sample to the laboratory for inspection. During the test, the collected clinical samples are first eluted and diluted, and then added to the reaction dish or the test strip for color development. The pH measurement requires direct contact with the secretions for color observation, while other reactions are washed.
  • De-dilution can not be unified operation, at the same time after dilution, the detection is more indirect, it will reduce the sensitivity, the operation steps are more, the accuracy is poor, for the bedside doctor, there are multiple operation steps and judgment indicators, very time-consuming and cumbersome, for the experiment For the room, it is transferred to the laboratory for experimental operation, and the transfer time in the middle is long, which is easy to degrade and change the enzyme and other substances, affecting the authenticity and accuracy of the result.
  • Bacterial vaginosis in gynecology The vaginosis, BV) test is an example.
  • Bacterial vaginosis is one of the most common vaginal infections in women of childbearing age. The infection rate is 30-50%, the incidence rate is 10-20%, and the number of patients is much higher than that of vaginal trichomoniasis, mold, etc. And easy to relapse. In addition, about 50% of patients with bacterial vaginosis have been diagnosed and treated for lack of clinical symptoms.
  • the Amsel standard and the Nugent standard are internationally recognized as the two so-called "gold standard" for the diagnosis of bacterial vaginosis.
  • the Amsel standard can be diagnosed as bacterial vaginosis by having three of the following four indicators: (1) homogenous, thin, white secretions in the vagina; (2) vaginal pH > 4.5; (3) amine test Positive; (4) clue cells are positive.
  • the Nugent standard diagnoses BV by classifying and integrating different morphological flora in vaginal secretions.
  • the Amsel standard has the following shortcomings: 1.
  • the pH value is not convenient to measure.
  • Olfactory test staff is poorly compliant and objective, 4.
  • Cue cells are indirect evidence; Nugent standards also require smear, Gram staining, and then identify, count, and integrate individual bacterial species under oil mirrors, which are highly empirical, cumbersome, and time consuming.
  • the invention solves the technical problem of time-consuming, cumbersome and poor accuracy in detecting clinical liquid samples, and provides a biochip capable of directly detecting changes of microbial spectrum or biochemical substances in clinical liquid samples, which is fast, simple and objective. Strong and accurate, eliminating the disadvantages of multi-step operations such as collection, elution, dilution, and dropping, the results can be obtained directly at the bedside.
  • a biochip comprising a substrate and a film, the substrate being provided with a detecting unit, the film covering a surface of the detecting unit of the substrate.
  • the detecting unit may be a reaction spot formed by drying after adsorbing or combining different reaction reagents on the surface of the substrate.
  • the detecting unit may also be a detecting crucible fixed on the surface of the substrate, which is formed by adsorbing or combining different reaction reagents and drying the reaction carrier.
  • the detecting unit may also be a reaction spot on the film-form reaction carrier formed by adsorbing or combining different reaction reagents on a film-like reaction carrier, and the film-like reaction carrier is placed on the substrate.
  • the detecting unit may further be a reaction trap disposed on the substrate, and the bottom of the reaction well adsorbs or combines different reaction reagents and is dried.
  • the film is a semi-permeable membrane having micropores, which allows the liquid to be tested to rapidly enter and react with the reagent, while preventing the reflux of the liquid to be tested and preventing the reagent from oozing out of the film.
  • the substrate is an optically transparent material.
  • the biochip is provided with a barcode or a two-dimensional code.
  • biochip application for preparing a product for detecting a clinical fluid sample.
  • the clinical fluid sample includes vaginal secretions, oral mucus, gastrointestinal secretions, blood, body fluids, pleural effusion, or urine.
  • the biochip of the invention can directly detect the change of the microbial spectrum or the biochemical substance in the clinical liquid sample, eliminates the collection of the clinical liquid sample transfer test, elutes and dilutes, and then adds the elution droplet to the reaction test paper or reacts. These steps are developed in the reaction well of the plate, which is fast, convenient, direct and objective.
  • the tester can take the biochip out of the inspection site or from the liquid to be tested and directly put it into the photoelectric reading.
  • the instrument obtains the test result, which is suitable for bedside operation of the hospital outpatient doctor or the family use of the patient. It is the instantiation and bedside of the test medicine.
  • the embodiment of the care testing, POCT The embodiment of the care testing, POCT).
  • 1 to 4 are schematic views showing the structure of a biochip of the present invention.
  • the biochip 100 of the present invention comprises a substrate 110 and a film 120, wherein the substrate 110 is square or rectangular, and may also be circular or elliptical, having an area of 1 to 800 square millimeters and a thickness of 0.5 to 10 mm, the optimized area is 30 to 300 mm 2 , and the thickness is 1 to 3 mm; the substrate 110 is colorless and optically transparent.
  • a recess 111 is formed in the middle of both sides of the substrate 110, and the front sides of the substrate 110 are Each of the 1/3 central portions has a knife-shaped card slot, and one surface of the substrate is subjected to surface treatment to physically adsorb or chemically bond specific detection reagents and is dried to form different detection units 130.
  • the reagents of the detection unit 130 may be various reactions. Molecules such as antigens, antibodies, DNA, RNA, enzymes, enzyme substrates, biochemical reagents or mixtures thereof can react with specific molecules at the detection site, and different reagent groups are adsorbed or bonded to the surface of the substrate 110 as different detection units. On the treated surface, a reaction spot arranged in an array is formed, and then a film 120 is coated on the surface of the substrate having the reaction spot. The film 120 has micropores and liquid. And biological macromolecules can pass through, the film preferably having a unidirectional liquid, allowing the liquid to quickly enter into contact and reacts with the reagent, while preventing reflux of liquid reagent can prevent leaking to the outside the film.
  • Molecules such as antigens, antibodies, DNA, RNA, enzymes, enzyme substrates, biochemical reagents or mixtures thereof can react with specific molecules at the detection site, and different reagent groups are adsorbed or bonded to the surface of the
  • the detection reagent is adsorbed or bound to the reaction carrier 140, dried to form a detection unit, and different detection units are fixed on the substrate 110 to form an array arrangement.
  • a substrate 120 is coated on the substrate 110 having the detection flaw;
  • the reaction carrier 140 may be made of a polymer material such as synthetic resin particles, silica gel particles or natural fibers such as filter paper, paper fibers or inorganic materials such as alumina particles, nanometers. Gold particles, gold and metal oxide composite particles; various organic or inorganic mesh materials can also be used.
  • the detection reagent is adsorbed or bound to the membranous reaction carrier 150, dried to form the detection unit 151, and the different detection units 151 are arrayed on the membranous reaction carrier.
  • the reaction spot is a substrate 110, and the reaction carrier 150 is covered with a film 120; the film reaction carrier 150 can be made of a polymer material such as a nitrocellulose film, a non-woven fabric or a natural fiber material such as filter paper or inorganic. Fiber materials such as glass fibers.
  • the reaction wells 160 are arranged in an array on the substrate 110, the substrate 110 is colorless and optically transparent, and the depth of the reaction well 160 is 0.1 to 10 mm. Optimized is 0.5 to 3 mm, the number of reaction wells 160 is determined by the number of indicators to be detected, and the reaction well 160 may be of any shape; the bottom surface of the reaction well 160 serves as a detection window for detecting color reactions; and the reaction well 160 has adsorption.
  • a reaction carrier which is combined with a detection reagent and dried may be a polymer material such as synthetic resin particles, silica gel particles, or the like, or a natural fiber such as filter paper, cellulose, or inorganic microparticles or fiber materials such as alumina particles, nanometers. Gold particles, gold and metal oxide composite particles, glass fibers, etc.; various organic or inorganic mesh materials can also be used; the reaction carrier is prepared by mixing cellulose, super absorbent resin, and reagents to form a test slurry. The test slurry is then injected into the reaction trap and dried. The substrate 110 is covered with a film 120 while covering the reaction well opening.
  • reaction carrier 140 is superposed in multiple layers, each layer being impregnated with a different reaction reagent.
  • reaction carrier 150 is superposed in multiple layers, each layer being impregnated with a different reaction reagent.
  • the bottom surface of the reaction well 160 may be directly adsorbed or chemically bonded with a reagent and dried by chemical or physical treatment.
  • an optical fiber is formed on the bottom of the reaction tank on the substrate 110 for detecting the color change of the detecting unit.
  • the film covering the surface of the substrate can be replaced by a filter film bag, that is, the substrate and the reaction unit are wrapped by the filter film bag, and the filter film containing the biochip is taken out from the detection site. After the bag, the filter bag is peeled off.
  • the working principle of the biochip of the present invention is based on the characteristics of a color change or the luminescence characteristic of a reaction system after certain specific enzymatic reactions, immune reactions or chemical reactions, and the presence or amount of a specific measured molecule or ion is perceived.
  • Change the amount of which is related to the depth of the color or the brightness of the light, and the molecule or ion may represent some damage, imbalance of metabolism, the presence or amount of certain bacteria, or reflect changes in the pH of the environment.
  • Commonly known techniques include dry chemical method, enzyme-linked immunosorbent assay, colloidal gold, immunoluminescence, electrochemiluminescence, fluorescent labeling, etc. The so-called dry chemical method occurs after the application of certain biochemical reactions or enzymatic chemical reactions.
  • the principle of judging whether or not the molecule to be tested exists and the amount thereof is determined by color change, and the reagent required for the reaction is impregnated or attached to the filter paper and dried, and the sample is added to the filter paper to observe the color change during the detection.
  • the reagent required for the reaction is impregnated or attached to the filter paper and dried, and the sample is added to the filter paper to observe the color change during the detection.
  • the chemotactic action of polymorphonuclear leukocytes accumulates in the vagina and releases a large amount of leukocyte esterase.
  • the activity of the esterase released by polymorphonuclear leukocytes can be detected, which can reflect vaginal secretion.
  • Cleanliness, degree of vaginal mucosal damage, is an important indicator for the diagnosis of vaginitis; according to leukocyte esterase hydrolysis of 5-bromo-4-chloro-3-indolyl acetate, release of bromoguanidine, the latter It is blue in the presence of oxygen, and has a chemical reaction property in which the color depth is proportional to the activity of leukocyte esterase.
  • the reaction carrier such as filter paper impregnated substrate 5-bromo-4-chloro-3-indole acetate is dried.
  • the above chemical reaction can occur when the vaginal fluid is encountered, and the activity of the leukocyte esterase can be known by measuring the depth of the blue color to determine the degree of vaginal secretion cleanliness, vaginal mucosal damage or inflammation.
  • the activity of the leukocyte esterase can be known by measuring the depth of the blue color to determine the degree of vaginal secretion cleanliness, vaginal mucosal damage or inflammation.
  • the invention can integrate the detection indicators such as the detection targets into different kinds of detection units according to different detection purposes to form a biochip, and the biochip is attached to the vaginal wall during the detection, the vaginal liquid penetrates into the biochip, and the collection is directly detected.
  • the object has a variety of lesion information, and then the biochip is placed in the optoelectronic reader to assist in the diagnosis of vaginal diseases by reading and comprehensively analyzing the biochip information.
  • the biochemical reagents on the reaction carrier and the different amounts and kinds of reaction carriers can be replaced according to different diagnostic objects, so the biochip of the invention can be used not only for vaginal microenvironment detection, but also for oral cavity, gastrointestinal tract, and extraction. Blood, body fluids, pleural effusion, urine or dilutions of these liquids.
  • the biochip 100 of the present invention comprises a substrate 110 and a film 120 having an area of about 80 mm 2 and a thickness of about 1 mm.
  • the substrate 110 is made of colorless optically transparent polymethyl methacrylate ( PMMA); one side of the substrate 110 is subjected to surface treatment such as chrome gelatin, and is physically adsorbed or chemically combined with a specific detection reagent and dried to form a pH value, hydrogen peroxide, leukocyte esterase, sialidase, hydrazine.
  • PMMA colorless optically transparent polymethyl methacrylate
  • each detection unit 130 includes a detection membrane block and a blank control membrane block, each membrane block having a length and width of about 1.2 mm, respectively, to 3
  • the array of ⁇ 4 is arranged on the substrate, and the hydrophilic nonwoven fabric having micropores is covered thereon, and is pressed and fixed to the substrate.
  • the biochip 100 of the present invention comprises a substrate 110, a reaction carrier 140 impregnating the detection reagent, and a film 120 having an area of about 130 mm 2 and a thickness of about 2 mm.
  • the substrate 110 is colored and transparent.
  • Methyl methacrylate ( PMMA) one surface of the substrate 110 is treated with chrome gelatin; the synthetic resin particles are used as a reaction carrier impregnation reagent to prepare pH, hydrogen peroxide, leukocyte esterase, sialidase, proline aminopeptidase, respectively.
  • each detecting unit comprising a detecting membrane block and a blank control membrane block, each of the membrane blocks having a length and a width of about 1.5 mm, arranged on the substrate in a 3 ⁇ 4 array, Further, a hydrophilic nonwoven fabric having micropores is covered thereon, and is pressed and fixed to the substrate. There is a recess 111 in the middle of both side faces of the substrate.
  • the biochip 100 of the present invention is composed of a substrate 110, a reaction carrier 150 impregnating a detection reagent, and a film 120 having an area of about 144 square millimeters and a thickness of about 2 mm.
  • the substrate 110 is colored and optically transparent.
  • Methyl methacrylate PMMA
  • one plate of substrate 110 is treated with chrome gelatin; filter paper is used as reaction carrier impregnation reagent to prepare pH, hydrogen peroxide, leukocyte esterase, sialidase, proline aminopeptidase, acetyl
  • An glucosaminidase detecting unit each detecting unit comprising a detecting membrane block and a blank control membrane block, each of the membrane blocks having a length and a width of about 1.8 mm, arranged on the substrate in a 3 ⁇ 4 array, and then The upper surface is covered with a hydrophilic nonwoven fabric having micropores, and is pressed and fixed to the substrate.
  • the biochip 100 of the present invention comprises a substrate 110, a reaction carrier and a film 120.
  • the substrate 110 has an area of about 192 square millimeters and a thickness of about 2 millimeters.
  • the substrate 110 is made of colorless optically transparent polydimethylsiloxane. Alkane (PDMS); substrate 110 has an array of reaction wells 160; each reaction well 160 has a length and width of about 1.5 mm and a depth of about 1 mm, arranged in a 4 x 6 array on the substrate.
  • PDMS polydimethylsiloxane
  • Pulp, super absorbent resin (SAP), and reagents are mixed to prepare pH, hydrogen peroxide, leukocyte esterase, sialidase, proline aminopeptidase, acetylglucosaminidase, nitrite, Oxidase, trichomonas-specific hydrolyzed protein, glucosaminidase activity, polyamine, heme detection slurry, and then injected into the reaction well 160, dried; and then covered with microporous hydrophilic non-woven fabric , pressed and fixed with the substrate.
  • a recess 111 is formed in the middle of both sides of the substrate.

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Abstract

一种生物芯片，包括基板和薄膜，所属基板上设有检测单元，所述薄膜覆盖在该基板的检测单元表面。上述生物芯片在制备检测临床液体样本的产品中的应用。该生物芯片可以直接检测临床液体样本中微生物谱或生化物质的变化情况，快速、简便、客观性强、准确度高，应用前景非常广阔。

Description

生物芯片及其应用
技术领域
本发明涉及临床诊断医疗器械技术领域，尤其涉及一种生物芯片及其应用。
背景技术
目前，临床上检测阴道分泌物、口腔粘液、胃肠道分泌物、抽出的血液、体液、胸腹水、或尿液等，通常都需要先采集阴道分泌物、口腔粘液、或胃肠道分泌物等临床液体样本，再将样本拿到实验室进行检验操作。检验时，一般先将采集的临床样本进行洗脱稀释，然后滴加到反应皿或试纸条上经显色等过程，pH值测定要求直接接触分泌物显色观察，而其他的反应经过洗脱稀释，不能统一操作，同时经过稀释后检测比较间接，也会降低灵敏度，操作步骤多、准确性差，对于床边医生来说，有多个操作步骤和判断指标，非常费时和繁琐，对于实验室来说，移送到实验室进行实验操作，中间的移送交接时间较长，对于酶等物质容易降解发生变化，影响结果的真实性和准确度。
以妇科的细菌性阴道病(bacterial vaginosis，BV)检测为例。细菌性阴道病是育龄妇女最常见的阴道感染性疾病之一，感染率在30-50%，发病率在10-20%，患病人数远远高于阴道滴虫、霉菌等的感染人数，且易复发。另外，约50%的细菌性阴道病患者因无明显临床症状而被贻误诊断和治疗。目前，Amsel标准和Nugent标准是国际上公认的两大诊断细菌性阴道病的所谓“金标准”。Amsel标准为同时拥有以下四项指标中的三项即可以诊断为细菌性阴道病：（1）阴道有匀质、稀薄、白色分泌物；（2）阴道pH值>4.5；（3）胺试验阳性；（4）线索细胞阳性。Nugent标准是通过对阴道分泌物中不同形态菌群分类和积分来诊断BV。
在临床实践中，Amsel标准存在以下不足：1．pH值测定不方便，2．阴道分泌物性状判断主观性强，3．嗅试验工作人员顺应性和客观性差，4．线索细胞是间接证据；Nugent标准也存在需要涂片、革兰氏染色，然后在油镜下对单个细菌的种类进行鉴别、计数和积分，专业经验性强，繁琐、费时。
随着对BV发病机制的不断深入了解，产生了以检测BV患者阴道分泌物中所特有的微生物的代谢产物为基础的诊断方法并出现了一些商业试剂盒，比如中国知识产权局公开号为CN201247244的“妇科多项目干化学联合检测试纸条”和公开号为 CN101975852A的“一种细菌性阴道病快速检测装置、检测试纸条及检测方法”，这些方法较上述方法客观且适于批量操作，但还需将标本拿到实验室检验操作，要肉眼判断颜色变化，都需要用试子采集阴道分泌物、洗脱稀释，然后滴加到反应皿或试纸条上显色等过程。操作步骤多、准确性差，非常费时和繁琐。
因此目前急需一种能快速、准确、全面地检测临床液体标本的医疗器材。
发明内容
本发明要解决目前检测临床液体标本费时、繁琐、准确性差的技术问题，提供一种生物芯片，该生物芯片可以直接检测临床液体样本中微生物谱或生化物质的变化情况，快速、简便、客观性强、准确度高，免去了采集、洗脱、稀释、滴加等多步骤操作的缺点，可以在床边直接得到结果。
此外，还需要提供一种上述生物芯片的应用。
为了解决上述技术问题，本发明通过如下技术方案实现:
在本发明的一个方面，提供了一种生物芯片，包括基板和薄膜，所述基板上设有检测单元，所述薄膜覆盖在该基板的检测单元表面。
所述检测单元可以是通过在基板的表面吸附或结合不同的反应试剂后经干燥而形成的反应斑点。
所述检测单元也可以是固定在基板表面的检测垛，该检测垛是由反应载体吸附或结合不同反应试剂后经干燥而形成。
所述检测单元也可以是通过在膜状反应载体上吸附或结合不同的反应试剂后经干燥而形成的位于该膜状反应载体上的反应斑点，该膜状反应载体置于所述基板上。
所述检测单元还可以是设于基板上的反应阱，该反应阱内底部吸附或结合有不同反应试剂并经干燥。
优选的，所述薄膜是具有微孔的半透膜，能够让待测液体迅速进入与试剂接触并发生反应，同时既能防止待测液体反流又能防止试剂渗出到薄膜外面。
优选的，所述基板是光学透明材质。
优选的，所述生物芯片上设有条形码或二维码。
在本发明的另一方面，还提供了一种生物芯片的应用，用于制备检测临床液体样本的产品。
所述临床液体样本包括阴道分泌物、口腔粘液、胃肠道分泌物、血液、体液、胸腹水、或尿液。
本发明的生物芯片，可以直接检测临床液体标本中微生物谱或生化物质的变化情况，免去了采集临床液体标本移送检验科，洗脱稀释，然后将洗脱液滴加到反应试纸上或反应板的反应孔内显色这些操作步骤，快速、便捷、直接、客观性强，配合使用光电阅读仪，检测者可以将生物芯片从检查部位或从被测液体中拿出后直接放入光电阅读仪获得检测结果，适合于医院门诊医生床边操作或患者家庭使用，是检验医学即时化和床边化（point of care testing，POCT）的体现。
附图说明
下面结合附图和具体实施方式对本发明作进一步详细的说明。
图1至图4是本发明生物芯片的结构示意图。
其中：100、生物芯片； 110、基板； 120、薄膜； 130、检测单元； 140、反应载体； 150、膜状反应载体； 151、膜状反应载体上的检测单元； 160、反应阱； 111、基板两侧面中部凹陷。
具体实施方式
如图1所示，本发明的生物芯片100，包括基板110和薄膜120，其中，基板110是方形或长方形，也可以是圆形或椭圆形，面积是1至800平方毫米，厚度是0.5至10毫米，优化后面积是30至300平方毫米，厚度是1至3毫米；基板110是无色光学透明的，优选的，在基板110的两侧面中部有一个凹陷111，基板110两侧面的前1/3中部各有一个刀形卡槽，该基板一个面经过表面处理可以物理吸附或化学结合特定检测试剂并经干燥，形成不同的检测单元130，检测单元130的这些试剂可以是各种反应分子比如抗原、抗体、DNA、RNA、酶、酶底物、生化试剂或其混合物，可以与检测部位的特定分子发生反应，不同的试剂组作为不同的检测单元吸附或结合在基板110的经过表面处理的那一个面上，形成阵列排布的反应斑点，然后再在有反应斑点的基板面覆一层薄膜120，这种薄膜120有微孔，液体和生物大分子能够透过，该薄膜最好具有单向导液性，能够让液体迅速进入与试剂接触并发生反应，既能防止液体反流又能防止试剂渗出到薄膜外面。
作为本发明生物芯片的另一种体现方式如图2所示，检测试剂吸附或结合在反应载体140上，经干燥形成检测单元，不同的检测单元固定在基板110上，形成阵列排布的检测垛，然后再在有检测垛的基板110面覆一层薄膜120；反应载体140可以采用高分子材料比如合成树脂颗粒、硅胶颗粒或者天然纤维比如滤纸、纸纤维或者无机材料比如氧化铝颗粒、纳米金颗粒、金和金属氧化物复合颗粒；也可以采用各种有机或无机的网眼状材料。
作为本发明生物芯片的另一种体现方式如图3所示，检测试剂吸附或结合在膜状反应载体150上，经干燥形成检测单元151，不同的检测单元151在膜状反应载体上阵列排布成反应斑点，反应载体150下面是基板110，反应载体150上面覆盖一层薄膜120；膜状反应载体150可以采用高分子材料比如硝酸纤维素膜、无纺布或者天然纤维材料比如滤纸或者无机纤维材料比如玻璃纤维等。
作为本发明生物芯片的另一种体现方式如图4所示，基板110上有反应阱160呈阵列状排布，基板110是无色光学透明的，反应阱160的深度是0.1至10毫米，优化的是0.5至3毫米，反应阱160的个数由需要检测的指标多少而定，反应阱160可以是任何形状；反应阱160的底面作为探测颜色反应的探测窗；反应阱160内有吸附或结合检测试剂并经干燥的反应载体，该反应载体可以采用高分子材料比如合成树脂颗粒、硅胶颗粒等，或者天然纤维比如滤纸、纤维素，或者无机微颗粒或纤维材料比如氧化铝颗粒、纳米金颗粒、金和金属氧化物复合颗粒、玻璃纤维等；也可以采用各种有机或无机的网眼状材料；该反应载体的制作过程是将纤维素、高吸水性树脂、试剂混合形成检测浆，然后再将该检测浆注入所述反应阱，干燥处理。基板110上面覆盖一层薄膜120，同时盖住反应阱口。
作为本发明的改进，反应载体140是多层叠加的，每层分别浸渍有不同的反应试剂。
作为本发明的改进，反应载体150是多层叠加的，每层分别浸渍有不同的反应试剂。
作为本发明的改进，反应阱160底面经过化学或物理处理可以直接吸附或由化学键结合有反应试剂并经干燥。
作为本发明的改进，基板110上反应槽底部有光纤，用来探测检测单元的颜色变化。
作为本发明的改进，生物芯片上有条形码或二维码。
作为本发明的另一种体现方式是所述覆盖在基板表面的薄膜可以用过滤膜袋取代，即用过滤膜袋将基板和反应单元包裹，待从检测部位拿出包有生物芯片的过滤膜袋后，将过滤膜袋剥除。
本发明生物芯片的工作原理是基于某些特定的酶促反应、免疫反应或化学反应后反应体系能够发生颜色变化的特性或发光的特性，感知某种特定的被测分子或离子存在或量的变化，其量的多少与颜色的深浅或光的亮度相关，而这种分子或离子可能代表机体某种损伤、代谢的失衡、某种细菌的存在或量的变化，或者反映环境的酸碱度的变化；通常公知的技术方法有干化学法、酶联免疫法、胶体金、免疫发光法、电化学发光、荧光标记法等，所谓干化学法是应用某些生物化学反应或者酶促化学反应后发生颜色变化而判断被测分子是否存在及其量的多少的原理，将反应所需要的试剂浸渍吸附或联接在滤纸上经干燥处理，检测时将样本加在滤纸上观察颜色变化。比如阴道内发生炎症反应时，由于多形核白细胞的趋化作用在阴道内聚集而大量释放白细胞酯酶，在这种炎症情况下检测多形核白细胞释放的酯酶的活性，可以反应阴道分泌物清洁度、阴道粘膜损伤的程度，是辅助诊断阴道炎的一个重要指标；根据白细胞酯酶水解5-溴-4-氯-3-吲哚乙酸盐，释放出溴吲哚基，后者在氧存在的条件下呈蓝色，呈色深度与白细胞酯酶活性成正比的化学反应性质，将反应载体比如滤纸浸渍底物5-溴-4-氯-3-吲哚乙酸盐经干燥处理，当遇到阴道液时可以发生上述化学反应，通过测量蓝色的深浅度可以得知白细胞酯酶的活性进而判断阴道分泌物清洁度、阴道粘膜损伤或炎症的程度。同样，通过检测乳酸、氧化酶、滴虫特异性水解蛋白、pH值、过氧化氢浓度、唾液酸苷酶活性、脯氨酸氨基肽酶活性、氨基葡萄糖苷酶活性、多胺、亚硝酸盐、亚铁血红素等信息来综合判断阴道内哪些细菌是优势菌群、有无菌群的失衡、酸碱度高低、有无滴虫、霉菌或大肠杆菌感染等微生态情况。本发明可以将诸如此类的检测指标根据检测目的不同制成不同种类的检测单元集成到一块基板上制成生物芯片，检测时将生物芯片贴在阴道壁上，阴道液渗入生物芯片，直接采集被检对象多种病变信息，然后将生物芯片放入光电阅读仪，通过阅读和综合分析生物芯片信息来辅助诊断阴道疾病。
本发明根据不同的诊断对象可以更换反应载体上的生化试剂以及不同数量和种类的反应载体，因此本发明的生物芯片不仅可以用于阴道微环境检测，还可用于口腔、胃肠道、抽出的血液、体液、胸腹水、尿液或这些液体的稀释液等。
实施例1
如图1所示，本发明生物芯片100由基板110和薄膜120构成，基板110面积约是80平方毫米，厚度约是1毫米，基板110用无色光学透明聚甲基丙烯酸甲酯( PMMA)制成；基板110一个面经过表面处理如铬明胶处理后，通过物理吸附或化学结合特定检测试剂并经干燥，形成检测pH值、过氧化氢、白细胞酯酶、唾液酸苷酶、脯氨酸氨基肽酶、乙酰氨基葡萄糖苷酶等不同的检测单元130，每一种检测单元130包括一个检测膜块和一个空白对照膜块，每一个膜块长宽各约是1.2毫米，以3×4的阵列排布于基板上，再在上面覆盖具有微孔的亲水性无纺布，与基板压合固定。基板的两侧面中部有一个凹陷111。
实施例2
如图2所示，本发明生物芯片100由基板110、浸渍检测试剂的反应载体140和薄膜120构成，基板110面积约是130平方毫米，厚度约是2毫米，基板110用无色光学透明聚甲基丙烯酸甲酯( PMMA)制成；基板110的一个板面经过铬明胶处理；用合成树脂颗粒作为反应载体浸渍试剂分别制成pH值、过氧化氢、白细胞酯酶、唾液酸苷酶、脯氨酸氨基肽酶、乙酰氨基葡萄糖苷酶检测单元，每一种检测单元包括一个检测膜块和一个空白对照膜块，每一个膜块长宽各约是1.5毫米，以3×4的阵列排布于基板上，再在上面覆盖具有微孔的亲水性无纺布，与基板压合固定。基板的两侧面中部有一个凹陷111。
实施例3
如图3所示，本发明生物芯片100由基板110、浸渍检测试剂的反应载体150和薄膜120构成，基板110面积约是144平方毫米，厚度约是2毫米，基板110用无色光学透明聚甲基丙烯酸甲酯( PMMA)制成；基板110的一个板面经过铬明胶处理；用滤纸作为反应载体浸渍试剂分别制成pH值、过氧化氢、白细胞酯酶、唾液酸苷酶、脯氨酸氨基肽酶、乙酰氨基葡萄糖苷酶检测单元，每一种检测单元包括一个检测膜块和一个空白对照膜块，每一个膜块长宽各约是1.8毫米，以3×4的阵列排布于基板上，再在上面覆盖具有微孔的亲水性无纺布，与基板压合固定。基板的两侧面中部有一个凹陷111。
实施例4
如图4所示，本发明生物芯片100由基板110、反应载体和薄膜120构成，基板110面积约为192平方毫米，厚度约是2毫米，基板110用无色光学透明聚二甲基硅氧烷（PDMS）制成；基板110上有反应阱160阵列状排布；每一个反应阱160的长宽各约是1.5毫米，深度约是1毫米，以4×6的阵列排布于基板上；将纸浆、高吸水性树脂(SAP)、试剂混合分别制成pH值、过氧化氢、白细胞酯酶、唾液酸苷酶、脯氨酸氨基肽酶、乙酰氨基葡萄糖苷酶、亚硝酸盐、氧化酶、滴虫特异性水解蛋白、氨基葡萄糖苷酶活性、多胺、亚铁血红素检测浆，然后注入反应阱160内，干燥处理；再在上面覆盖有微孔的亲水性无纺布，与基板压合固定。所述基板的两侧面中部有一个凹陷111。
以上所述实施例仅表达了本发明的实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (10)

  1. 1.一种生物芯片，其特征在于，包括基板和薄膜，所述基板上设有检测单元，所述薄膜覆盖在该基板的检测单元表面。
  2. 2.根据权利要求1所述的生物芯片，其特征在于，所述薄膜是具有微孔的半透膜。
  3. 3.根据权利要求1所述的生物芯片，其特征在于，所述检测单元是通过在基板的表面吸附或结合不同的反应试剂后经干燥而形成的反应斑点。
  4. 4.根据权利要求1所述的生物芯片，其特征在于，所述检测单元是固定在基板表面的检测垛，该检测垛是由反应载体吸附或结合不同反应试剂后经干燥而形成。
  5. 5.根据权利要求1所述的生物芯片，其特征在于，所述检测单元是通过在膜状反应载体上吸附或结合不同的反应试剂后经干燥而形成的位于该膜状反应载体上的反应斑点，该膜状反应载体置于所述基板上。
  6. 6.根据权利要求1所述的生物芯片，其特征在于，所述检测单元是设于基板上的反应阱，该反应阱内底部吸附或结合有不同反应试剂并经干燥。
  7. 7.根据权利要求1至6中任一项所述的生物芯片，其特征在于，所述基板是光学透明材质。
  8. 8.根据权利要求7所述的生物芯片，其特征在于，所述生物芯片上设有条形码或二维码。
  9. 9.权利要求1所述生物芯片的应用，其特征在于，用于制备检测临床液体样本的产品。
  10. 10.根据权利要求9所述的应用，其特征在于，所述临床液体样本包括阴道分泌物、口腔粘液、胃肠道分泌物、血液、体液、胸腹水、或尿液。
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