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Gold-labeled test paper strip for quick diagnosis of hemorrhagic fever with renal syndrome

A technology for renal syndrome and hemorrhagic fever, applied in the direction of biological testing, preparation of test samples, sugar derivatives, etc., to achieve the effects of reducing costs, reducing fatality rates, and high sensitivity

Inactive Publication Date: 2006-07-12
FUJIAN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Retrieval of relevant materials including Chinese patents shows that there is no use of genetic engineering methods to obtain highly expressed and highly active truncated NP recombinant antigens for the diagnosis of hemorrhagic fever with renal syndrome and the simultaneous detection of IgM and IgG antibodies in the serum of HFRS patients Related reports on HFRS gold standard rapid diagnostic test strips

Method used

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  • Gold-labeled test paper strip for quick diagnosis of hemorrhagic fever with renal syndrome
  • Gold-labeled test paper strip for quick diagnosis of hemorrhagic fever with renal syndrome

Examples

Experimental program
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Effect test

Embodiment 1

[0052] 1. Preparation of recombinant antigen

[0053] 1. Hantavirus strain A used in this study 537 It is a strain directly isolated from the lung tissue of Apodemus syringae captured in the HFRS epidemic area of ​​Zhouning County, Fujian Province in 1985 by using Vero E6 cells;

[0054] 2. Take the virus infection supernatant, and extract viral RNA with a Viral RNA Mini Kit (QIAGEN company) extraction kit.

[0055] 3. Use P 14 Reverse transcription primer and S 1 / S 2 PCR amplification primers from Hantavirus A 537 A full-length S gene of 1696 bp was amplified in the strain genome. The RT-PCR reaction system is as follows:

[0056] reactive components

Volume (μl)

5×AMV buffer

dNTP (10mmol / L)

p14 (10μmol / L)

RNase inhibitor

AMV reverse transcriptase

RNA template

h 2 o

4

1

1

0.5

1

5

7.5

[0057] Reverse transcription reaction conditions: 10 minutes at room temperature, 1 hour of rever...

Embodiment 2

[0110] Preparation of HFRS gold standard rapid diagnostic test strips

[0111] (1) Get 1000ml of 0.03% tetrachloroauric acid, heat to boiling, add 25ml of 1% trisodium citrate solution, continue to boil for 5 minutes, after the color of the colloidal gold solution changes from blue to purple to red, cool naturally for later use;

[0112] (2) Take 1ml of colloidal gold solution, add 0.2M K 2 CO 3 10μl, 50μg of purified antigen, stir well and let stand for 1h;

[0113] (3) Add 20% bovine serum albumin 25μl, 20% PEG 20,000 15μl, centrifuge at 8,000r / min for 8min, and discard the supernatant;

[0114] (4) Suspend the precipitate in 1ml of basic buffer, which is the gold-labeled antigen solution;

[0115] (5) Label mouse IgG in the same way;

[0116] (6) After the two gold-labeled antigen solutions are mixed, the glass fiber paper is fully infiltrated, and placed in a lyophilizer to lyophilize;

[0117] (7) Goat anti-mouse IgG monoclonal antibody 3.0mg / ml, goat anti-human μ ch...

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Abstract

The invention discloses a recombination antigen for HFRS diagnosis and its rapid GIA diagnosis indicator paper opposite to shortened NP recombination antigen e112 of Hantaan virus by gene clone technique and simultaneous-detected IgM and IgG antigens in patient serum. The said test paper overcomes the limitation of current HFRS diagnosis method, benefit to early-stage diagnosis, reduces illness death rate, does not requires special device nor professional staff to meet more detection need, and cuts cost greatly.

Description

technical field [0001] The invention relates to an antigen for disease diagnosis, in particular to a recombinant antigen for diagnosis of hemorrhagic fever with renal syndrome and a rapid gold standard diagnostic test strip thereof. Background technique [0002] Hantaviruses (HV) are widespread and harmful, and have become a global public health problem. Hemorrhagic fever with renal syndrome (HFRS) caused by HV infection is a group of acute infectious diseases with fever, bleeding and renal damage as the main clinical manifestations. The epidemic areas are distributed in 32 countries and regions in Asia, Europe, Africa, and the Americas, but they are mainly prevalent in my country and South Korea in Asia, followed by Russia, Finland, and the former Yugoslavia in Europe. There are fewer cases in Africa and the Americas; the source of the epidemic The land covers more than 70 countries on five continents of the world. In China, HFRS is the most harmful viral infectious diseas...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531G01N33/52C07H21/00C07K16/00G01N1/28
Inventor 严延生吴守丽
Owner FUJIAN CENT FOR DISEASE CONTROL & PREVENTION
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