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Porcine parvovirus assay kit and application thereof

A parvovirus and kit technology, which is applied in the detection/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of cumbersome operation, expensive equipment, high cost, etc., and achieve high specificity, simple and convenient operation, The effect of high sensitivity

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

However, PCR technology also has its shortcomings. The main problems are high cost, strong technicality, cumbersome operation, expensive equipment, and high sample detection cost, so it is not suitable for primary diagnosis and large-scale epidemiological investigation.

Method used

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  • Porcine parvovirus assay kit and application thereof
  • Porcine parvovirus assay kit and application thereof
  • Porcine parvovirus assay kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, the preparation of porcine parvovirus detection kit

[0048] 1. Synthesis of primers

[0049] Artificially synthesize the following 3 pairs of primers:

[0050] F3: 5'-CAAGCAACAATGGCTAGCTATATG-3' (sequence 2 of the sequence listing);

[0051] B3: 5'-GTTGGTGTTGTTGGCTCGC-3' (sequence 3 of the sequence listing);

[0052] FIP: 5'-GTATTTATTGGGGTTTGCATTTTTTTATCATTGGGGAAATGTACCTGAT-3' (SEQ ID NO: 4 of the Sequence Listing);

[0053] BIP: 5'-ACATCAGTGAAAACTTCGCCAGCTTTTGCTAAATCCAGGTCCTCCTGT-3' (SEQ ID NO: 5 of the Sequence Listing);

[0054] LF: 5'-GCTCCTCCCATTTTTCTGAC-3' (sequence 6 of the sequence listing);

[0055] LB: 5'-ACAACAACTACGCAGCAACTC-3' (SEQ ID NO: 7 of the Sequence Listing).

[0056] 2. Preparation of sample tube

[0057] 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.

[0058] 3. Preparat...

Embodiment 2

[0065] Embodiment 2, the preparation of porcine parvovirus detection kit

[0066] 1. Synthesis of primers

[0067] Same as Step 1 of Example 1.

[0068] 2. Preparation of reaction tube

[0069] Same as Step 2 of Example 1.

[0070] 3. Preparation of positive control

[0071] Same as Step 3 of Example 1.

[0072] 4. Preparation of LAMP reaction solution

[0073] A liquid is the same as the A liquid of embodiment 1.

[0074] 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 14 pmol / μl.

[0075] 5. Assembling the kit

[0076] Same as Step 5 of Example 1.

Embodiment 3

[0077] Embodiment 3, the preparation of porcine parvovirus detection kit

[0078] 1. Synthesis of primers

[0079] Same as Step 1 of Example 1.

[0080] 2. Preparation of reaction tube

[0081] Same as Step 2 of Example 1.

[0082] 3. Preparation of positive control

[0083] Same as Step 3 of Example 1.

[0084] 4. Preparation of LAMP reaction solution

[0085] A liquid is the same as the A liquid of embodiment 1.

[0086]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 10pmol / μl.

[0087] 5. Assembling the kit

[0088] Same as Step 3 of Example 1.

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Abstract

The invention discloses a porcine parvovirus assay kit and application thereof. The kit provided by the invention comprises three pairs of primers, namely an inside primer pair, an outside primer pair and an annular primer pair combined with an NS1 gene in genome DNA (GenBank Accession Number NC-001718) of a porcine parvovirus. The porcine parvovirus assay kit of the invention has the advantage of high assay sensitivity and is simple and convenient for operation, can detect ten copied target DNAs at minimum and is particularly suitable for clinical medicinal detection and the detection of porcine parvovirus probably polluting foods in the basic level yield.

Description

technical field [0001] The invention relates to a porcine parvovirus detection kit and application thereof. Background technique [0002] Porcine parvovirus (Porcine parvovirus, PPV) is one of the important pathogens that cause reproductive disorders in pigs. It is mainly characterized by abortion, infertility, stillbirth, mummified fetuses and weak offspring in sows. In addition, PPV is also associated with non-suppurative myocarditis, PPV can aggravate multisystemic wasting syndrome (PMWS) in weaned piglets caused by porcine circovirus (PCV). In recent years, with the development of the pig industry, large-scale intensive pig farms have appeared one after another, and the occurrence of the disease is on the rise, causing huge economic losses to the pig industry and seriously affecting the development of the pig industry. [0003] In 1966, Mary and Mahnel first isolated the virus from swine fever virus tissue and confirmed its pathogenic role. Since then, it has been repo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 吴文学黄书林李佳禾
Owner CHINA AGRI UNIV
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