Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of porcine leocircle virus type 2 ring-mediated isothermal amplification kit and its application

A ring-mediated isothermal, micro-ring virus technology, applied in the field of microbial detection, can solve the problems of laboratory contamination, false positive results, unsuitable for grassroots and on-site detection, etc., and achieves the effect of simple interpretation and easy operation.

Active Publication Date: 2019-12-27
GUANGXI VETERINARY RES INST
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some deficiencies in the application of these technologies and they have high requirements for hardware. Agarose gel electrophoresis is still required to determine the results, which is likely to cause laboratory pollution and lead to false positive results, and it takes a long time. Suitable for grassroots and field testing

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of porcine leocircle virus type 2 ring-mediated isothermal amplification kit and its application
  • A kind of porcine leocircle virus type 2 ring-mediated isothermal amplification kit and its application
  • A kind of porcine leocircle virus type 2 ring-mediated isothermal amplification kit and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The specificity result of embodiment 1 LAMP detection method

[0082] LAMP amplification was carried out on 1 strain of porcine leovirus type 2, 7 strains of control virus and water control, the results were as follows figure 1 As shown, the turbidity rising curve appeared in the reaction tube of porcine leiocircular virus type 2 in about 18 minutes, which was a positive result, and the curves of the 7 strains of control virus reaction tubes and the water control reaction tube showed no amplification, which was a negative result.

Embodiment 2

[0083] Example 2 Sensitivity results of LAMP detection method

[0084] The starting concentration of porcine leovirus type 2 raw DNA was 1.40×10 2 ng / μL, after 10-fold serial dilution, carry out LAMP and PCR amplification, the results are as follows figure 2 with image 3 As shown, the result shows that the detection limit of the LAMP method of the present invention is about 1.40 × 10 -8 ng / μL, while the detection limit of conventional PCR method is 1.40×10 -7 ng / μL.

Embodiment 3

[0085] Example 3 Fluorescence visualization detection results of LAMP detection method

[0086] According to the optimized conditions monitored by the turbidimeter, fluorescent dyes were added to the reactor, and after 60 minutes of reaction at 63°C, observed under ultraviolet light, Figure 4 In order to observe the results, the left tube is the reaction with the porcine leovirus type 2 genome DNA as a template, which is a positive result, and the right tube is a negative control, which is a negative result. The test results show that the established LAMP method can be used conveniently at the grassroots level. You only need to use the kit with the LAMP primer designed by this method, add the sample, and use an inexpensive water bath to keep it at 63°C for 60 minutes to quickly observe the results without the need Open the lid to avoid contamination.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a porcine torque teno sus virus type 2 loop-mediated isothermal amplification kit and application thereof. The kit comprises an LAMP primer, a 2*reaction buffer solution, a BstDNA (BstDeoxyribonucleic Acid) polymerase, a fluorescent visual inspection reagent, ultra-pure water and a porcine torque teno sus virus type 2 DNA template, wherein the LAMP primer comprises outer primers F3 and B3, inner primers FIP and BIP and loop primers LF and LB. The application of the porcine torque teno sus virus type 2 loop-mediated isothermal amplification kit comprises the following steps: detecting a porcine torque teno sus virus type 2 diseased tissue sample by using the kit and performing porcine torque teno sus virus type 1 pathogen qualitative research. Specificity detection and sensitivity detection verify that an LAMP detection method provided by the invention can monitor the reaction in real time, quantitatively detect the copy number of a porcine torque teno sus virus type 1 and quickly and accurately obtain detection results, thereby bringing convenience to simple, convenient, quick and reliable detection of the porcine torque teno sus virus type 2.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a fast, visualized and real-time quantitative detection of porcine letrocyclovirus type 2 loop-mediated isothermal amplification kit and its application. Background technique [0002] TTSuV (Torque teno sus virus, TTSuV) is a non-enveloped, single-strand negative-strand circular DNA virus. According to its nucleotide sequence differences, TTSuV can be divided into two subtypes, TTSuV1 and TTSuV2. Among them, TTSuV1 was classified into the genus Iota-torquevirus, and TTSuV2 was classified into the genus Kappatorque-virus. Letrocyclovirus was first discovered in the serum of post-transfusion hepatitis patients by Japanese scholar Nishizawa et al. in 1997. Its host range is very wide. At present, this virus has widely infected non-human primates, domestic and wild animals, and humans. Letrocyclovirus has a high infection rate in the population. According to the epidemio...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12R1/93
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2561/113C12Q2545/113C12Q2563/107
Inventor 陈忠伟何颖段群棚周英宁赵武秦毅斌卢冰霞李斌梁家幸杨思仪蒋冬福苏乾莲
Owner GUANGXI VETERINARY RES INST