Ternary PCR detection primers and detection method for goat pathogenic bacteria

A technology for detecting primers and detection methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of long time-consuming, low sensitivity, etc., and achieve the advantages of simple operation, guaranteed specificity, and shortened detection time Effect

Inactive Publication Date: 2015-07-01
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the identification of goat-causing pathogens includes isolation and culture and electron microscope observation, which is a time-consuming process with high requirements for operation. Low sensitivity of diagnostic methods such as electrophoresis, reverse passive hemagglutination test, latex agglutination test, single expansion hemolysis test or ELISA
Most of the PCR det

Method used

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  • Ternary PCR detection primers and detection method for goat pathogenic bacteria
  • Ternary PCR detection primers and detection method for goat pathogenic bacteria
  • Ternary PCR detection primers and detection method for goat pathogenic bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Taking 3 strains of goat pox virus, 3 strains of Clostridium wilchii and 3 strains of Mycoplasma capricum isolated clinically, the DNA templates were prepared by the existing DNA extraction method.

[0025] The goat pox virus primer sequence designed and synthesized according to the goat pox virus P32 gene is:

[0026] P32-F: 5'-CTCATTGGTGTTCGGATT-3'

[0027] P32-R: 5'-ATGGCAGATATCCCATTA-3'.

[0028] According to the Clostridium wilchii α toxin cpa gene design and synthesis of the Clostridium wilchii primer sequence is:

[0029] cpa-F: 5'-TAATGTTACTGCCGTTGATAGCG-3'

[0030] cpa-R: 5'-CATAATCCCAATCATCCCAACTA-3'.

[0031] According to the Mycoplasma capricolum 16S rRNA gene design and synthesis of the Mycoplasma capricolum primer sequence is:

[0032] 16S rRNA-F: 5'-ACTATGAGATGGGGATGCGGC-3'

[0033] 16S rRNA-R: 5'-GGACTTTAACCTCAAACTTGC-3'.

[0034] The concentration of the upper and lower primers of the goatpox virus is 5.4 μmol / L, the concentration of the upper and...

Embodiment 2

[0039] Triple PCR reaction conditions and reaction steps are the same as in Example 1, except that the mixed DNA template of goat pox virus, Clostridium wilchii and Mycoplasma capricum is used as the test object.

[0040] Using the mixed DNA templates of 3 strains of goat pox virus, Clostridium wilchii and Mycoplasma capricum as the test object, the DNA templates were prepared by the existing DNA extraction method.

[0041] The goat pox virus primer sequence designed and synthesized according to the goat pox virus P32 gene is:

[0042] P32-F: 5'-CTCATTGGTGTTCGGATT-3'

[0043] P32-R: 5'-ATGGCAGATATCCCATTA-3'.

[0044] According to the Clostridium wilchii α toxin cpa gene design and synthesis of the Clostridium wilchii primer sequence is:

[0045] cpa-F: 5'-TAATGTTACTGCCGTTGATAGCG-3'

[0046] cpa-R: 5'-CATAATCCCAATCATCCCAACTA-3'.

[0047] According to the Mycoplasma capricolum 16S rRNA gene design and synthesis of the Mycoplasma capricolum primer sequence is:

[0048] 16S rRN...

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Abstract

The invention discloses ternary PCR detection primers and detection method for goat pathogenic bacteria, wherein goatpox virus, clostridium welchii of goat and caprine mycoplasmas can be detected at the same time. The invention relates to three pairs of PCR primers which specifically amplify gene sequences of goatpox virus, clostridium welchii of goat and caprine mycoplasmas, respectively; the three pairs of primers can be amplified in a same PCR reaction tube at the same time to realize multiple detection, so that the detection time is greatly shortened and the method is simple and convenient to operate. Deign of the specific primers ensures high conservation and specificity of the primers and avoids non-complementary pairing or cross-species amplification between the primers and a DNA template. The method disclosed by the invention is small in time consumption, high in efficiency and high in sensitivity.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a goat pathogen triple PCR detection primer and a detection method capable of simultaneously detecting goat pox virus, Clostridium wilchii and Mycoplasma capricum. technical background [0002] Goat pox is an acute, febrile, contact infectious disease caused by goat pox virus (Goatpox virus, GTPV). Papule-pustular pox occurs on the skin. Clostridium welchii, also known as Clostridium perfringens (Clostridium perfringens), belongs to saprophytic anaerobic spore-causing bacteria, and is the main pathogen causing "sudden death syndrome" in goats. The bacteria are divided into A, B, C, D, E 5 different bacterial types, α-toxin is a necrotic and lethal toxin produced jointly by them. Some people think that α-toxin is the main pathogenic factor of "sudden death syndrome" in livestock. Mycoplasmal pneumonia of sheep and goats, also known as sheep infectious pleuropneumonia...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04C12N15/11C12R1/93C12R1/145C12R1/35
CPCC12Q1/70C12Q1/6844C12Q1/689C12Q2600/16
Inventor 王豪举范文萍秦溢杨红军徐丽敏
Owner SOUTHWEST UNIVERSITY
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