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A kind of method suitable for the elisa of toxoplasma gondii

A Toxoplasma, labeling technology, applied in the field of ELISA applicable to Toxoplasma, can solve problems such as unclear key molecules

Active Publication Date: 2019-11-05
SHENYANG AGRI UNIV
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  • Claims
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Problems solved by technology

[0003] The key molecule of Toxoplasma gondii causing host pathogenicity is still unclear

Method used

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  • A kind of method suitable for the elisa of toxoplasma gondii
  • A kind of method suitable for the elisa of toxoplasma gondii
  • A kind of method suitable for the elisa of toxoplasma gondii

Examples

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Embodiment 1

[0056] Embodiment 1: the establishment of pig Toxoplasma gondii ELISA detection method

[0057] 1. Extraction of total RNA from Toxoplasma gondii ME49 strain

[0058] a. Take out the worm sediment from -80°C, add 1 mL of sterilized PBS to wash, and centrifuge for 10 min at 2500 rpm.

[0059] b. After centrifugation and washing, discard the supernatant PBS, add 1mL Trizol to the precipitate, and repeatedly pipette to ensure that the worms can be completely broken and lysed.

[0060] c. Add 200 μL of chloroform to 1 mL of Trizol, shake for 15 s with a shaker, and let stand on ice for 3 min. d. After standing still, place in a centrifuge at 4°C, centrifuge for 15 minutes at 12000×g, draw 400 μL of the supernatant and place it in a new sterile EP tube.

[0061] e. Add the same volume (400 μL) of isopropanol, invert and mix for 1 min, and let stand on ice for 10 min.

[0062] f. After standing still, centrifuge in a centrifuge at 4°C for 10 minutes at 12000g / min, discard the sup...

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Abstract

The invention provides a method of ELISA applicable to toxoplasma gondii. The method is characterized by comprising the following steps of using TGME49_223140 His label recombinant protein and GST label recombinant protein to detect pig serum; conducting double enzyme digestion on pEASY Simple Blunt plasmid containing a target fragment and using the same restriction enzyme to conduct double enzymedigestion on expression carriers pET-28a and pGEX4T-1; converting the recombinant expression carriers pET-28a and pGEX4T-1 into engineering bacteria E.coilBL21(DE3) and E.coil BL21 respectively to express and purify protein; using the purified TGME49_223140 His label recombinant protein and GST label recombinant protein as covering protein respectively and building an ELISA detection method.

Description

technical field [0001] The invention relates to the acquisition, expression and purification of the TGME49_223140 gene of toxoplasma gondii and the establishment of an ELISA method. Background technique [0002] Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii (Toxoplasma gondii), which has a worldwide distribution; it is seriously harmful to people and animals with immune insufficiency and immunosuppression. [0003] The key molecules of Toxoplasma gondii causing host pathogenicity are still unclear. [0004] Toxoplasmosis threatens human health and the development of animal husbandry. Contents of the invention [0005] Purpose of the present invention: to establish a sensitive and effective ELISA detection method for Toxoplasma gondii. [0006] The technical scheme provided by the invention is as follows: [0007] 1 nucleotide sequence [0008] (1) Acquire nucleic acid sequence from TOXODB (http: / / toxodb.org / toxo / ) database. [0009] (2) The...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/70C12N15/66G01N33/96G01N33/68
CPCC12N15/66C12N15/70G01N33/68G01N33/96
Inventor 陈启军李烨馨姜宁杨娜桑晓宇冯颖
Owner SHENYANG AGRI UNIV
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