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

A primer set, kit and application for quantitative detection of Toxoplasma gondii

A quantitative detection, Toxoplasma gondii technology, applied in biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., can solve false negative, false negative results or low quantitative value, loss and other problems, to achieve specific Strong, rigorous design, accurate and objective results

Active Publication Date: 2022-07-08
JILIN UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have shown that clinical specimens such as serum, whole blood, tissue, and cerebrospinal fluid contain a large number of unknown components, which will inhibit the PCR reaction, and the residual reagents in the nucleic acid extraction process will also inhibit PCR amplification, resulting in false positives. Negative results or low quantitative values
On the other hand, loss during nucleic acid extraction can also cause false negative results

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 primer set, kit and application for quantitative detection of Toxoplasma gondii
  • A primer set, kit and application for quantitative detection of Toxoplasma gondii
  • A primer set, kit and application for quantitative detection of Toxoplasma gondii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] This embodiment provides a primer set for quantitative detection of Toxoplasma gondii, and its design method is as follows:

[0022] After the sequence comparison of multiple strains of Toxoplasma gondii, it was found that the Toxoplasma gondii L358 gene is located in regions 63-82, 338-354 and other regions, and the bases are well conserved, so primers were designed for these segments. Among them, for the Toxoplasma gondii L358 gene, a segment of this gene that is common to all species and has the smallest inter-species variation is selected, and the conserved part of the sequence is compared in NCBI. This segment is shared by multiple Toxoplasma gondii, and the homology is greater than 70% , meets the genus-specific detection standard, and its sequence is shown in SEQ ID NO: 3 in the sequence listing.

[0023] When designing, try to ensure that the GC content of the selected region is between 40%-60%, the amplification length is between 100bp-400bp, and the Tm value o...

Embodiment 2

[0027] This embodiment provides a kit for quantitatively detecting Toxoplasma gondii, which includes a PCR reaction solution, a primer set, a positive control substance and a negative control substance.

[0028] Wherein, the PCR reaction solution can use commercially available 2*SYBR Green MIX, which includes dNTP, fluorescent dye, Taq polymerase and reaction buffer.

[0029] The primer set includes a forward amplification primer and a reverse amplification primer. The nucleotide sequence of the forward amplification primer is as shown in SEQ ID NO: 1 in the sequence table, specifically 5'-GAGAAAGCGAAACCTTCCTG-3'; The nucleotide sequence is shown in SEQ ID NO: 2 of the sequence table, specifically 5'-CGACACACTCGCATGCATG-3'; in addition, the concentrations of the forward amplification primer and the reverse amplification primer are both 10 μmol / L.

[0030] The negative control was deionized water; the positive control was a plasmid cloned from a specific fragment of the Toxopla...

Embodiment 3

[0032] This embodiment provides a kit for quantitatively detecting Toxoplasma gondii, which includes a PCR reaction solution, a primer set, a positive control substance and a negative control substance.

[0033] Wherein, the PCR reaction solution can use commercially available 2*SYBR Green MIX, which includes dNTP, fluorescent dye, Taq polymerase and reaction buffer.

[0034] The primer set includes a forward amplification primer and a reverse amplification primer. The nucleotide sequence of the forward amplification primer is as shown in SEQ ID NO: 1 in the sequence table, specifically 5'-GAGAAAGCGAAACCTTCCTG-3'; The nucleotide sequence is shown in SEQ ID NO: 2 of the sequence table, specifically 5'-CGACACACTCGCATGCATG-3'; in addition, the concentrations of the forward amplification primer and the reverse amplification primer are both 12 μmol / L.

[0035] The negative control was deionized water; the positive control was a plasmid cloned from a specific fragment of the Toxopla...

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 is applicable to the field of biotechnology, and provides a primer set, kit and application for quantitative detection of Toxoplasma gondii, the primer set includes a forward amplification primer and a reverse amplification primer; the core of the forward amplification primer is The nucleotide sequence is shown in SEQ ID NO: 1 in the sequence listing; the nucleotide sequence of the reverse amplification primer is shown in SEQ ID NO: 2 in the sequence listing. The primer set for quantitative detection of Toxoplasma gondii provided by the present invention is designed and optimized by selecting targeted Toxoplasma gondii gene fragments, the design is rigorous, the operation is simple and fast, the sensitivity is high, and the specificity is strong, and can be applied to human infection of Toxoplasma gondii The detection of pathogenic nucleic acids in blood or cerebrospinal fluid samples is accurate and objective.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer set, a kit and application for quantitatively detecting Toxoplasma gondii. Background technique [0002] Toxoplasma Gondii is an intracellular parasite, also known as three corpses. It is parasitic in the cells, flows with the blood, and reaches all parts of the body, destroying the brain, heart, and fundus of the eyes, resulting in decreased immunity and various diseases. It is an obligate intracellular parasite, Coccidia, Eucalyptus, Isosporaceae, Toxoplasma. The life cycle requires two hosts. The intermediate hosts include reptiles, fish, insects, birds, mammals and other animals and humans, and the final hosts include cats and felines. Studies have found that most people are carriers of Toxoplasma gondii, and they form worm immunity, and it is difficult to develop symptoms of initial infection after being infected. However, in HIV patients, due to the weak...

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): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2545/114Y02A50/30
Inventor 张文艳郑柏松刘新李兆龙桓晨高文英王虹
Owner JILIN UNIV