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Detection method and detection reagent kit for nephrotic syndrome and hemorrhagic fever

A detection kit and technology for renal syndrome, applied in the fields of molecular biology, immunology, and infectious diseases, can solve problems such as not being disclosed

Inactive Publication Date: 2002-12-04
上海市南汇县南华医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no published gold drop method and kit for IgG

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Preparation of Hantavirus Antigen

[0053] The hamtavirus strain 76-118 of the wild mouse type was inoculated into gerbil kidney cells, and after conduction culture at 35-37° C. for 7-10 days according to a conventional method, the cell culture was collected.

[0054] After ultrasonic treatment, the cells were roughly centrifuged at 3000rpm for 30 minutes to remove impurities such as cell membrane fragments. The supernatant was taken and centrifuged at 40,000 rpm for 2 hours to precipitate the hantavirus antigen. Discard the supernatant and take the pellet. After the precipitate was dissolved, the protein content in the solution was determined. If the protein content is low, it can be centrifuged again to make the final concentration of Hantavirus antigen 1-2MG / ML.

Embodiment 2

[0056] Preparation of colloidal gold

[0057] Add 100 ml of deionized double-distilled water to a 250 ml Erlenmeyer flask, add 4 ml of 1% sodium citrate, and quickly add 1 ml of 1% chloroauric acid after boiling. Boiling was continued for 15 minutes and after cooling the pH was adjusted to 8.5 with 0.1M potassium carbonate.

Embodiment 3

[0059] Preparation of Colloidal Gold-Goat Anti-IgG Antibody Conjugate

[0060] (a) Preparation of goat anti-IgG antibody The goat anti-IgG antibody purchased from Shanghai Institute of Biological Products was purified by ammonium sulfate for use.

[0061] (b) Stability test Since the potency of various biological products purchased (or prepared by conventional methods) fluctuates, in order to improve repeatability, it is advisable to continue stability tests on each batch of antibodies to determine the best antibody Add amount.

[0062] Take a batch of goat anti-IgG antibody solution with an antibody concentration of about 1 mg / ml as an example. In 1 ml of the colloidal gold solution prepared in Example 2, add 0 microliters, 2 microliters, 4 microliters, 6 microliters, 8 microliters, 10 microliters, 12 microliters respectively, and mix at room temperature After homogenization, let stand at room temperature for 10 minutes. Add 0.1 ml of 10% sodium chloride to each tube, mix ...

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Abstract

The invention provides a method for detecting hemorrhagic fever with renal syndrome by a gold drop method and a detection kit. In this method, the anti-Hantavirus IgG antibody in the serum is used as the detection object, and it includes the steps of: (a) coating the purified Hantavirus nucleoprotein on a nitrocellulose membrane to form a coating in the sampling area. A nitrocellulose detection membrane with hantavirus nucleoprotein; (b) using non-human anti-human IgG antibody and colloidal gold as raw materials to prepare a conjugate of colloidal gold-anti-human IgG antibody; (c) the Serum sample is added dropwise on the spotting area of ​​the detection membrane obtained in step (a); (d) washing the spotting area of ​​the detection membrane of step (c); (e) colloidal gold-antihuman The conjugate of the IgG antibody is dropped on the spotting area; (f) washing the spotting area of ​​the detection membrane in step (e). The method and kit can detect hemorrhagic fever with renal syndrome rapidly, correctly and conveniently.

Description

technical field [0001] The present invention relates to the fields of epidemiology, molecular biology and immunology. More specifically, the present invention relates to a detection method and a detection kit for hemorrhagic fever with renal syndrome. Background technique [0002] Hemorrhagic fever with renal syndrome (HFRS) is an acute infectious disease in humans caused by certain viruses in the genus Hantavirus of the Kenyaviridae family. The mortality rate is high after the onset, so early correct diagnosis is very necessary. In my country, hemorrhagic fever with renal syndrome (HFRS) is the most harmful infectious disease except viral hepatitis. The scope of the epidemic area of ​​this disease involves Europe, Asia, Africa and other parts of the world, and the number of cases in my country has no decreasing trend. In the Pudong area of ​​Shanghai, due to the increase of the foreign population, the incidence of HFRS is mostly high. Therefore, the early rapid diagnosi...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/544
Inventor 储峰季青严润民王霞明
Owner 上海市南汇县南华医院
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