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Polymerase chain reaction (PCR) kit for detecting sheep clostridium perfringens

A detection kit, the technology of Clostridium wilchii in sheep, which is applied in the field of kits, achieves the effect of simple detection method, good use effect and avoiding errors

Inactive Publication Date: 2011-11-16
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Now the market lacks a kit that can detect Clostridium wilchii in clinical samples simply, quickly, sensitively, accurately and reproducibly

Method used

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  • Polymerase chain reaction (PCR) kit for detecting sheep clostridium perfringens
  • Polymerase chain reaction (PCR) kit for detecting sheep clostridium perfringens
  • Polymerase chain reaction (PCR) kit for detecting sheep clostridium perfringens

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Effect test

Embodiment 1

[0054] Embodiment 1 of the present invention: Clostridium wilchii PCR detection kit, it comprises the PCR premix damping fluid of 250 μ L (PCR premix damping fluid is made of the MgCl of 3 mmol / μ L 2 , 500pmol / μL of dNTP, 25mmol / μL of Tris-HCl and 0.2U Taq DNA polymerase / μL, the pH value of which is pH8.3), 150μL of ultrapure water, 50μL of Marker DL2000, 40μL of 10μmol / L The upper primer (the sequence of the upper primer is LPF 5'-TAATGTTACTGCCGTTGATAGCG-3') and 40 μL of the lower primer with a concentration of 10 μmol / L (the sequence of the lower primer is 5'-CATAATCCCAATCATCCCAACTA-3'), 20 μL of ultrapure water as a negative control and 20 μL of standard Clostridium welchii PCR product was used as a positive control.

Embodiment 2

[0055] Embodiment 2 of the present invention: Clostridium welchii sheep PCR detection kit, it comprises the PCR premix damping fluid of 250 μ L (PCR premix damping fluid is made of the MgCl of 3 mmol / μ L 2 , 500pmol / μL of dNTP, 25mmol / μL of Tris-HCl and 0.5U Taq DNA polymerase / μL, the pH value of which is pH8.3), 150μL of ultrapure water, 50μL of Marker DL2000, 40μL of 9μmol / L The upper primer (the sequence of the upper primer is LPF 5'-TAATGTTACTGCCGTTGATAGCG-3') and 40 μL of the lower primer with a concentration of 9 μmol / L (the sequence of the lower primer is 5'-CATAATCCCAATCATCCCAACTA-3'), 20 μL of ultrapure water as a negative control and 20 μL of standard Clostridium welchii PCR product was used as a positive control.

Embodiment 3

[0056] Embodiment 3 of the present invention: Clostridium welchii PCR detection kit, it comprises the PCR premix damping fluid of 250 μ L (PCR premix damping fluid is made of the MgCl of 3 mmol / μ L 2 , 500pmol / μL of dNTP, 25mmol / μL of Tris-HCl and 0.4U Taq DNA polymerase / μL, the pH value of which is pH8.3), 150μL of ultrapure water, 50μL of Marker DL2000, 40μL of 11μmol / L The upper primer (the sequence of the upper primer is LPF 5'-TAATGTTACTGCCGTTGATAGCG-3') and 40 μL of the lower primer with a concentration of 11 μmol / L (the sequence of the lower primer is 5'-CATAATCCCAATCATCCCAACTA-3'), 20 μL of ultrapure water as a negative control and 20 μL of standard Clostridium welchii PCR product was used as a positive control.

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Abstract

The invention discloses a polymerase chain reaction (PCR) kit for detecting sheep clostridium perfringens. The kit comprises 250mu L of PCR premix buffer solution, 150mu L of ultrapure water, 50mu L of marker DL2000, 40mu L of 10mu mol / L upper primer, 40mu L of 10mu mol / L lower primer, 20mu L of negative control and 20mu L of positive control. By optimizing PCR conditions and replacing labor-consuming and time-consuming biochemical experiment and serological monitoring with the conventional PCR method, the PCR kit for detecting sheep clostridium perfringens disease is developed; and the kit can detect the sheep clostridium perfringens disease, can be used for the early and quick diagnosis of the sheep clostridium perfringens disease, has great significance for preventing and treating the sheep clostridium perfringens disease, can avoid errors caused by artificial judgment, shortens the detection time of the sheep clostridium perfringens disease, and has the characteristics of high speed, sensitivity and specificity and the like.

Description

technical field [0001] The invention relates to a kit, in particular to a PCR detection kit for Clostridium welchii. Background technique [0002] Clostridium welchii, also known as Clostridium perfringens ( Clostridium prerfringens ) belongs to saprophytic anaerobic spore-causing bacteria, and is the main pathogenic bacteria causing sudden death syndrome, necrotic enteritis, enterotoxemia and gas gangrene in livestock and poultry (W Muller, 1998). There are 5 different bacterial types of the bacteria, A, B, C, D, and E (Laetitia Petit et al., 1999). The pathogenicity is mainly related to the toxins they form. There are 12 types of toxins produced, and the α toxins are jointly produced by them A necrotic, lethal toxin considered by some to be a major cause of "sudden death syndrome" in livestock (Rood JI et al., 1991; Hale MI et al., 1999). Clostridium welchii disease in sheep caused by Clostridium welchii mainly occurs in sheep toxemia, including sheep sudden attack, shee...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 冉懋韬余波谭诗文艾玉萍徐景峨魏赐开
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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