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Quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and preparation method and quantitative determination method thereof

A quantitative detection and quantum dot technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of high consumption of reagents, complex detection process, long time required for analysis, etc., and achieve a wide range of applications.

Inactive Publication Date: 2014-12-10
CHENGDU LINGYU BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have many deficiencies: ELISA method to detect samples: ① The samples to be tested need centrifuge to separate the serum first, and need to use incubators, microplate readers, plate washing machines and other equipment; ② Only one indicator can be detected at a time; ③ The detection process is complex and time-consuming, and it takes at least 40-60 minutes to detect each indicator; ④The detection generally requires a certain number of samples to be carried out; ⑤On-site rapid detection cannot be realized
The analysis takes a long time, consumes a lot of reagents, the analysis throughput is low, and the labor load is large

Method used

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  • Quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and preparation method and quantitative determination method thereof
  • Quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and preparation method and quantitative determination method thereof
  • Quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and preparation method and quantitative determination method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: Quantum dot labeling test strip structure capable of quantitatively detecting multiple indicators of blood infectious diseases

[0048] combine figure 1 be explained. figure 1 Among them, the quantum dot labeled test strip capable of quantitatively detecting multi-indicators of blood infectious diseases includes a sample pad 1, a red blood cell filter membrane 2, a marker pad 3, an analysis membrane 7, and Absorbent pad6.

[0049] The marking pad 3 of the test strip is a glass fiber membrane. The analysis membrane 7 is a nitrocellulose membrane, a nylon membrane, or a nitrocellulose / acetate mixed membrane, on which there are a detection zone (ie T zone) 4 and a quality control zone (ie C zone) 5 . Described backing 8 is polyester or plastic plate.

[0050] The marker pad 3 is coated with CdSe / ZnS QD 523 Labeled HBsAg monoclonal antibody (i.e. CdSe / ZnS QD 523 -HBsAg Mc), CdSe / ZnS QD 616 Labeled HIVAg (i.e. CdSe / ZnS QD 616 -HIVAg), CdSe / ZnS QDs 56...

Embodiment 2

[0051] Embodiment 2: Preparation method of quantum dot-labeled test strip capable of quantitatively detecting multi-indicators of blood infectious diseases

[0052] The preparation method of described test strip comprises the steps:

[0053] A. Antibody / antigen related to quantum dot coupling:

[0054] a) Take water-soluble CdSe / ZnS QD 523 、CdSe / ZnS QD 616 、CdSe / ZnS QD 568 、CdSe / ZnS QD 457 Use PBS buffer to adjust pH=6-9, respectively add EDC (l-(3-dimethylaminopropyl)-3-ethylcarbodiamine hydrochloride) and NHS (N-hydroxythiosuccinic acid imine) activated at room temperature for 10-60min;

[0055] b) Add HBsAg Mc, HIVAg, HCVAg, TPAg and vortex shake respectively for 0.5-3h;

[0056] c) Add bovine serum albumin (BSA) respectively, and block the reaction in the dark for 0.5-2h;

[0057] d) Each product was purified by centrifugation;

[0058] e) The precipitates of each product were resuspended and dispersed in PBS buffer to obtain CdSe / ZnS QD 523 -HBsAg Mc,...

Embodiment 3

[0067] Example 3: Establishment of a standard curve for quantum dot-labeled test strips capable of quantitatively detecting multiple indicators of blood infectious diseases

[0068] （a） Using phosphate buffered solution (PBS) as the diluent, prepare 9 copies of HBsAg, HIVAb, HCVAb, and TPAb standard substances in series as shown in the table below.

[0069]

[0070] （b） The concentration of each standard substance in the table was detected by 10 quantum dot-labeled test strips under the same conditions with a detector for 10 times, and the T-band fluorescence intensity (OD t ) vs. C-band fluorescence intensity (OD c ), to get the mean and OD t / OD c ratio.

[0071] （c） Take the concentration of the standard series as the X-axis, and use OD t / OD c The ratio is used as the Y axis to obtain the standard curve corresponding to the fluorescence intensity and concentration. figure 2 .

[0072] The concentration of the standard series can also be used as the X-axis, ...

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Abstract

The invention belongs to the field of in-vitro diagnosis, and concretely relates to a quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and a preparation method and a quantitative determination method. A marking pad (3) of the test strip is coated with a quantum-dot marked mixture of HBsAg monoclonal antibody, HIVAg, HCVAg and TPAg, a T band (4) of an analysis membrane (7) is coated with a mixture of HBsAg monoclonal antibody, HIVAg, mouse anti-human antibody and TPAg, and a C band (5) of the analysis membrane (7) is coated with secondary antibody. The standard curves of all detected substances are stored and installed on the test strip by employing electron labels. The test strip employs a detector with signal detection function to read the standard curves stored in the electron labels and combines with the to-be-tested sample corresponding fluorescence intensity measured by the detector, so as to synchronously quantitatively determine the concentration of HBsAg, HIVAb, HCVAb and TPAb in the samples.

Description

Technical field [0001] The invention is a field of in vitro diagnostic, which involves a quantum dot tag test bar and its method that can quantitatively detect multiple indicators of blood infectious diseases. Background technique [0002] Hepatitis B, hepatitis C, AIDS, and syphilis are four common infectious diseases related to blood safety.The Ministry of Health of my country stipulates that blood donors must check the hepatitis B virus surface antigen (HBSAG), hepatitis C virus antibody (HCVAB), HIV antibody (HIVAB), syphilis spiral antibody (TPAB). All hospitals and blood products must also check the above 4. Poin blood infectious disease. [0003] At present, the above four blood infectious disease tests often use enzyme-linked immune adsorption methods (ELISA) and GOLD-IMMUNOCHROMATOGRAPHY ASSAY (GICA).There are many shortcomings in these methods: ELISA method detection sample: ① The samples to be checked need to be separated by a centrifugal machine first, and instruments...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533
CPCG01N33/558G01N33/543
Inventor 侯飞王春英马义才熊妮娅
Owner CHENGDU LINGYU BIOTECH
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