Kit for PCR (Polymerase Chain Reaction) detection of haemophilus paragallinarum

A technology of Haemophilus gallinosaurs and a detection kit, which is applied in the field of PCR detection kits for Haemophilus paragallinarum, can solve problems such as difficult cultivation, high nutritional requirements, and decreased egg production rate of laying hens, achieving low cost and high detection efficiency. High sensitivity and fast detection effect

Inactive Publication Date: 2014-03-26
SHANDONG BINZHOU BOLAIWEI BIOTECH
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Problems solved by technology

There are three serotypes of the bacteria, A, B, and C, which mainly cause poor growth of bred chickens, lower egg production rate of laying hens, and cause great economic losses to the chicken industry[2]
At present, the methods for diagnosing chicken infectious rhinitis mainly use traditional bacterial culture and biochemical identification of Haemophilus paragallinarum. Because Haemophilus paragallinarum has high

Method used

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  • Kit for PCR (Polymerase Chain Reaction) detection of haemophilus paragallinarum
  • Kit for PCR (Polymerase Chain Reaction) detection of haemophilus paragallinarum

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Embodiment 1

[0021] A Haemophilus paragallinarum PCR detection kit, characterized in that: said kit includes 20 μL of PCR buffer, 25 μL of ultrapure water, 5 μL of Marker DL2000, 3 μL of 15 pmol / μL upper primer, and 3 μL of 15 pmol / μL lower primer , 3 μL polymerase, 3 μL bromophenol blue, 15 μL TE buffer, 5 μL negative control, 5 μL positive control.

[0022] The above PCR buffer consists of 25 mmol / L MgCl 2 2μL, 5μL of 10×PCR buffer, 10mmol / L dNTPx2μL.

[0023] The sequence number of the upper primer is 5'-CCGCATAGAARRRRAAGA-3', and the sequence number of the lower primer is 5'-CCAACCTTCCTCACCACC-3'.

[0024] The above-mentioned negative control is the DNA fragment of non-Haemophilus paragallinarum.

[0025] The positive control above is the PCR product of the standard Haemophilus paragallinarum.

[0026] R in the above primers is a degenerate primer.

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Abstract

The invention relates to a kit and more particularly relates to a kit for PCR (Polymerase Chain Reaction) detection of haemophilus paragallinarum. The kit comprises 20 microliters of PCR buffer solution, 25 microliters of ultrapure water, 5 microliters of Marker DL2000, 3 microliters of 15pmol/microliter upper primers, 3 microliters of 15pmol/microliter lower primers, 3 microliters of polymerase, 3 microliters of bromophenol blue, 15 microliters of TE buffer solution, 5 microliters of negative control and 5 microliters of positive control. The kit has the beneficial effects that the kit has the advantage of rapid detection of a PCR detection technology and the kit is low in construction cost; the special primers are adopted so as to meet the detection of a PCR product of a gene segment which is 0.00953ng at the minimum; the detection sensitivity is high.

Description

technical field [0001] The invention relates to a kit, in particular to a PCR detection kit for Haemophilus paragallinarum. Background technique [0002] Chicken infectious coryza (Infectious coryza, IC) is an upper respiratory tract disease caused by Haemophilus paragallinarum (HPG), the most obvious symptom is the discharge of serous or mucous secretions from the nasal passages and sinuses , Facial edema and conjunctivitis, with obvious swelling of the wattle [1]. There are 3 serotypes of the bacteria, A, B, and C, which mainly cause poor growth of rearing chickens, lower egg production rate of laying hens, and cause great economic losses to the chicken industry [2]. At present, the methods for diagnosing chicken infectious rhinitis mainly use traditional bacterial culture and biochemical identification of Haemophilus paragallinarum. Because Haemophilus paragallinarum has high nutritional requirements, it is not easy to cultivate, and the bacteria is often mixed with othe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/04C12Q1/686C12Q2531/113
Inventor 郎景民马景霞吴洪才杨平郭显坡
Owner SHANDONG BINZHOU BOLAIWEI BIOTECH
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