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A primer set for diagnosing swine umbilical cord blood swine fever virus wild strain, a kit containing the primer set and its application

A technology of swine fever virus and umbilical cord blood, which is applied in the biological field, can solve the problems of insufficient sensitivity, limited template content of amplification results, false negatives, etc., and achieve the level of broadness and specificity, reliable technical support, and evaluation of protection Effect

Active Publication Date: 2020-04-03
湖南国测生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amplification result is limited by the template content of the detection object, and when the template amount is small, negative results are often obtained
In the field of porcine umbilical cord blood swine fever virus diagnosis, the amount of swine fever virus in porcine umbilical cord blood is usually very low, and the traditional conventional PCR detection technology usually has false negative results due to its insufficient sensitivity, which cannot meet the requirements

Method used

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  • A primer set for diagnosing swine umbilical cord blood swine fever virus wild strain, a kit containing the primer set and its application
  • A primer set for diagnosing swine umbilical cord blood swine fever virus wild strain, a kit containing the primer set and its application
  • A primer set for diagnosing swine umbilical cord blood swine fever virus wild strain, a kit containing the primer set and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The flow chart of kit of the present invention being used for diagnosing porcine umbilical cord blood swine fever virus wild strain infection is as follows figure 1 shown. Concretely comprise the following steps: (1) umbilical cord blood sample collection; (2) umbilical cord blood sample processing, extraction of nucleic acid RNA and reverse transcription of cDNA; (3) detection of swine fever virus E2 gene fragment in umbilical cord blood whole blood; (4) ) According to the test results, sort out and analyze the data of the classical swine fever virus E2 gene fragment; (5) evaluate the immune effect of the swine fever vaccine.

[0068] 1. Extraction of total RNA of CSFV in umbilical cord blood

[0069] (1) Collection of umbilical cord blood

[0070] Cord blood collection involves the following steps:

[0071] a. Take a clean penicillin bottle and cork, clean it, boil and sterilize it for 30 minutes, dry it and collect it for later use;

[0072] b. When the piglets a...

Embodiment 2

[0120] In the sow farm of manufacturer A, all the swine fever vaccines were immunized 21 days before production, and the sows were randomly divided into groups. Piglet umbilical cord blood of pigs, according to the method described in implementation 1, carries out the diagnosis of pig umbilical cord blood CSFV wild strain, evaluates the immune effect of CSF vaccine, and the situation of piglets infected with CSFV. The result is as figure 2 shown.

[0121] From figure 2 It can be seen that the positive control amplified a 272bp band, the negative control had no 272bp band amplification, and there was no specific 272bp band amplification in the umbilical cord blood of samples 1-6, indicating that the umbilical cord blood of samples 1-6 There is no swine fever virus, sows do not have swine fever virus detoxification and piglet infection, and the sows have a high level of health, which further indicates that the sows' swine fever vaccine immunity is effective and the immunizat...

Embodiment 3

[0123]In the sow farm of manufacturer B, all the swine fever vaccines were vaccinated 21 days before production, and the sows were randomly divided into groups. Piglet umbilical cord blood of pigs, according to the method described in implementation 1, carries out the diagnosis of pig umbilical cord blood CSFV wild strain, evaluates the immune effect of CSF vaccine, and the situation of piglets infected with CSFV. The result is as image 3 shown.

[0124] From image 3 It can be seen from the figure that the positive control amplified a 272bp band, the negative control had no 272bp band amplification, and there was no specific 272bp band amplification in the umbilical cord blood of samples 1-20, indicating that the umbilical cord blood of samples 1-20 There is no swine fever virus in the pig, there is no detoxification of swine fever virus and piglet infection in sows, and the health level of sows is high, which further shows that the immunization effect of swine fever vacci...

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Abstract

The invention discloses a swine umbilical cord blood swine fever virus wild strain diagnosing primer group, a kit containing the same and application of the swine umbilical cord blood swine fever virus wild strain diagnosing primer group. Swine umbilical cord blood swine fever virus wild strain diagnosing primer are shown in SEQ ID NO: 1-SEQ ID NO: 4. The swine umbilical cord blood swine fever virus wild strain diagnosing primer group can be applied to producing a swine umbilical cord blood swine fever virus wild strain diagnosing kit. The swine umbilical cord blood swine fever virus wild strain diagnosing kit is composed of an outer set of primers, an inner set of primers, negative control, positive control and 2*PCR (polymerase chain reaction) Mixture. The swine umbilical cord blood swine fever virus wild strain diagnosing primer and the kit containing the swine umbilical cord blood swine fever virus wild strain diagnosing primer group can specifically diagnose swine umbilical cord blood swine fever virus wild strains and distinguish swine fever virus vaccine strains from swine fever virus vaccine strains as well as swine fever virus from bovine viral diarrhea-mocosal virus and border disease virus, and meanwhile, are high in sensibility, good in repeatability and reliable in detection results. The detection method through the swine umbilical cord blood swine fever virus wild strain diagnosing primer group is simple in process, short in detecting time and simple in result evaluation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set for diagnosing swine umbilical cord blood swine fever virus wild strain, a kit containing the primer set and applications thereof. Background technique [0002] Classical swine fever virus (CSFV) is a member of the Flaviviridae family, Pestivirus genus. Swine fever virus is highly contagious and pathogenic, causing mass mortality in pigs and causing serious losses to the swine industry. Pigs may shed large amounts of virus from the time they are infected with CSFV until they show clinical symptoms. If the animal tolerates acute or subacute infection, it will produce antibodies or turn into a chronic infection case, which will continuously or intermittently shed the toxin throughout its life until death. If a pregnant sow is infected with CSFV, the virus can cross the placenta and infect the fetus, leading to miscarriage, mummification or stillbirth. If infection occur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2549/119C12Q2531/113
Inventor 喻正军李增强石建廖娟红
Owner 湖南国测生物科技有限公司
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