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Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease

Babesiosis equine and kit technology, applied in the field of biological detection, can solve the problems of time-consuming, poor specificity, expensive kits, etc., and achieve the effect of eradicating harm and ensuring healthy development

Inactive Publication Date: 2016-07-06
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention: the purpose of the present invention is to provide a kind of rapid diagnosis technology and kit of Babesia equinoides to solve the shortcomings of conventional detection kits such as expensive, poor specificity, time-consuming, etc.

Method used

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  • Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease
  • Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease
  • Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease

Examples

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Effect test

Embodiment 1

[0031] Example 1. Designing detection primers, fluorescent probes and obtaining local positive target gene fragments. The inventors designed specific primers based on the published Bc48 gene sequence as shown in SEQIDNo.1 and SEQIDNo.2. The designed specific primers used PCR Amplify the genome of Xinjiang local insect strains to obtain the target antigen gene. As a specific target antigen gene, its nucleotide sequence is shown in SEQ ID No. 4, and the target antigen gene sequence is used as a genetic marker for detecting Babesia equine Things. In addition, those skilled in the art should understand that specific fragments of this sequence can only be used as genetic markers for detecting Babesia equine disease.

[0032] Obtain the target antigen gene: (1) According to the published Bc48 gene sequence, design specific primers as shown in SEQ ID No. 1 and SEQ ID No. 2 and probes as shown in SEQ ID No. 3; (2) Use PCR technology to extract the genome of local strains Amplify the tar...

Embodiment 2

[0033] Example 2. Establishment of fluorescent PCR detection method for Babesia equine: (1) Use specific primers for routine amplification (see attached image 3 ); (2) Application of specific primers and probes for fluorescence amplification; (3) Optimization of fluorescence PCR amplification system; (4) Comparison of sensitivity, specificity and stability of fluorescence PCR methods (see attached Figure 4 , Attached Figure 5 , Attached figure 2 ); (5) Compare the fluorescent PCR method with other detection and diagnosis results to detect the coincidence rate.

Embodiment 3

[0034] Example 3. Development of a fluorescent PCR detection kit for Babesia horsetail

[0035] (1) Use of the kit: This kit is used to detect whether horses are suffering from Babesia equine disease. It is used to detect whether the blood samples of equine animals contain Babesia equine. This detection method can be used in epidemiology. Research and formulate measures to reduce the prevalence of Babesia horses in horse herds.

[0036] (2) The principle of the kit: This kit uses the MGB fluorescent PCR detection method. The TaqMan MGB probe is improved on the basis of the TaqMan probe. The biggest difference from the TaqMan probe is that its length can be shortened to 13bp. The distance between the fluorescent group and the quenching group is closer, and the quenching effect is better. The 3'end of the probe is labeled with a quenching group that does not emit light, and the fluorescence background is reduced. The resolution of the fluorescent tube is greatly improved. Reduce bac...

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Abstract

The invention belongs to the technical field of biological detection, and in particular relates to a babesia caballi disease detection kit based on a fluorescent quantitative PCR (polymerase chain reaction) scalar quantity positive control, which is used for clinical diagnosis and daily monitoring of equine animal infected babesia caballi disease. By designing detection primers as shown by SEQ ID No.1 and SEQ ID No.2 and a fluorescent probe as shown by SEQ ID No.3, combining absolute quantification analysis of a fluorescent quantitative PCR system, and optimizing a reaction system and cycling conditions, a fluorescent PCR detection method for the babesia caballi disease is established. Moreover, the fluorescent PCR detection kit for the babesia caballi disease based on a fluorescent quantitative PCR scalar quantity is established, and is suitable for clinical diagnosis and epidemiological monitoring of the babesia caballi disease.

Description

Technical field [0001] The present invention belongs to the technical field of biological detection, and specifically relates to a detection kit for Babesia equine infection established on the basis of a fluorescent quantitative PCR scalar positive control, which is used for clinical diagnosis and daily monitoring of equine animals infected with Babesiosis equine . Background technique [0002] Babesia equine disease is a blood protozoal disease caused by Babesiacaballi parasitic in the red blood cells of equine animals. The diseased animals are mostly manifested as anemia, jaundice, high fever, anorexia, weight loss, etc. symptom. This disease is particularly harmful to entry and exit animals, and is classified as a Class B disease by OIE. At present, the drugs circulating in the market for the treatment of Babesia horsetail are generally effective. Due to Xinjiang’s special geographical environment, vector ticks are widely distributed and the incidence of this disease is incr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 巴音查汗·盖力克张杨李永畅王振宝宋瑞其明·乌尔娜王美玲
Owner XINJIANG AGRI UNIV
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