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Toxoplasma gondii detection kit and application thereof

A technology of toxoplasma gondii and kit, which is applied in the field of toxoplasma gondii detection kit, can solve the problems of unsuitable grass-roots application, and achieve the effect of high sensitivity, high specificity, simple and convenient operation

Inactive Publication Date: 2011-04-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The molecular biology diagnosis method is mainly polymerase chain reaction (PCR), which is faster and more sensitive than the first two methods, but it needs to use a special PCR instrument, which is not suitable for grassroots applications

Method used

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  • Toxoplasma gondii detection kit and application thereof
  • Toxoplasma gondii detection kit and application thereof
  • Toxoplasma gondii detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, preparation of Toxoplasma gondii detection kit

[0047] 1. Synthesis of primers

[0048] Artificially synthesize the following 3 pairs of primers:

[0049] F3: 5'-TGGGAAGCGACGAGAGTC-3' (sequence 2 of the sequence listing);

[0050] B3: 5'-TGGATTCCTCTCCTACCCCT (sequence 3 of the sequence listing);

[0051] FIP: 5'-CCGGTGTCTCTTTTTCCACCCTTTTTTCGGAGAGGGAGAAGATGTTT-3' (SEQ ID NO: 4 of the SEQUENCE LISTING);

[0052] BIP: 5'-TCGTGGTGATGGCGGAGAATTTTCCTCCTCCCCTTCGTCCAA-3' (SEQ ID NO: 5 of the Sequence Listing);

[0053] LF: 5'-GGAAAAGCAGCCAAGCCG-3' (sequence 6 of the sequence listing);

[0054] LB: 5'-GAATTGAAGAGTGGAGAAGAGGGCG-3' (SEQ ID NO: 7 of the Sequence Listing).

[0055] 2. Preparation of sample tube

[0056] A commercially available 0.6ml centrifuge tube (requiring no nucleic acid contamination) was loaded into a 1.2mm diameter FTA card (Whatman, Cat No. WB120205) cut with a hole punch to obtain a sample tube.

[0057] 3. Preparation of positive con...

Embodiment 2

[0064] Embodiment 2, the preparation of Toxoplasma gondii detection kit

[0065] 1. Synthesis of primers

[0066] Same as Step 1 of Example 1.

[0067] 2. Preparation of reaction tube

[0068] Same as Step 2 of Example 1.

[0069] 3. Preparation of positive control

[0070] Same as Step 3 of Example 1.

[0071] 4. Preparation of LAMP reaction solution

[0072] A liquid is the same as the A liquid of embodiment 1.

[0073] Solution B is composed of solute and solvent; the solvent is double distilled water; the solute and its concentration are as follows: F3 is 3.6pmol / μl, B3 is 3.6pmol / μl, FIP is 28pmol / μl, BIP is 28pmol / μl, LF is 14pmol / μl, LB is 14pmol / μl.

[0074] 5. Assembling the kit

[0075] Same as Step 5 of Example 1.

Embodiment 3

[0076] Embodiment 3, preparation of Toxoplasma gondii detection kit

[0077] 1. Synthesis of primers

[0078] Same as Step 1 of Example 1.

[0079] 2. Preparation of reaction tube

[0080] Same as Step 2 of Example 1.

[0081] 3. Preparation of positive control

[0082] Same as Step 3 of Example 1.

[0083] 4. Preparation of LAMP reaction solution

[0084] A liquid is the same as the A liquid of embodiment 1.

[0085] Solution B is composed of solute and solvent; the solvent is double distilled water; the solute and its concentration are as follows: F3 is 2.4pmol / μl, B3 is 2.4pmol / μl, FIP is 20pmol / μl, BIP is 20pmol / μl, LF is 10pmol / μl, LB is 10 pmol / μl.

[0086] 5. Assembling the kit

[0087] Same as Step 3 of Example 1.

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Abstract

The invention discloses a toxoplasma gondii detection kit and application thereof. The kit provided in the invention comprises three pairs of primers, i.e. an inside primer pair, an outside primer pair and an annular primer pair which are combined with Gen Bank Accession Number AF146527 genes in a toxoplasma gondii genome. The toxoplasma gondii detection kit has high detection sensitivity, can beused for detecting 10 copied target DNAs at minimum, is simple and convenient for operation, and is especially suitable for regular checkup, base clinical medicine detection works and site immediate detection of a hoggery.

Description

technical field [0001] The invention relates to a toxoplasma detection kit and its application. Background technique [0002] Toxoplasmosis is a zoonotic protozoan disease caused by Toxoplasma gondii. Toxoplasma can infect a variety of animals: 45 mammals, 70 birds and 5 cold-blooded animals. Cats and felines are the final hosts of Toxoplasma gondii. In the final host, Toxoplasma gondii mainly invades the epithelial cells of the small intestine, causing intestinal diseases. In intermediate hosts (such as pigs, cattle, sheep, etc.), Toxoplasma gondii invades the intestinal wall, enters the cells of the mononuclear macrophage system through the blood or lymph, and spreads to tissues and organs throughout the body, such as the brain, lymph nodes, Liver, heart, lung, muscle, etc., cause multiple organ lesions in the whole body, and do great harm to the intermediate host. Pigs are the main susceptible animals to Toxoplasma gondii. According to related reports, the infection ra...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 吴文学索勋朱引洁邹杰郭盼盼
Owner CHINA AGRI UNIV
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