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Primer set and probe for detection of Chlamydia pneumoniae 98kda MOMP gene and application thereof

A Chlamydia pneumoniae and gene technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems that cannot meet the needs of early diagnosis of pathogens, and cannot meet the clinical needs of early active treatment, etc., to achieve Conducive to application and promotion, low detection limit, and the effect of reducing false positive rate

Active Publication Date: 2016-04-20
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because it takes a certain amount of time for pathogens to enter the body for replication and antigen stimulation to produce antibodies, this method still cannot meet the needs of early pathogen diagnosis, and thus cannot meet the clinical needs of early active treatment

Method used

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  • Primer set and probe for detection of Chlamydia pneumoniae 98kda MOMP gene and application thereof
  • Primer set and probe for detection of Chlamydia pneumoniae 98kda MOMP gene and application thereof
  • Primer set and probe for detection of Chlamydia pneumoniae 98kda MOMP gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] The screening of embodiment 1 primer and probe

[0117] 1. Design of primers and probes:

[0118] Use the NCBI database to collect the 98KDa major outer membrane protein (MOMP) gene sequence of Chlamydia pneumoniae, use the biological software BioEdit to analyze and compare, and find out the conserved regions that are highly conserved and meet the detection sequence length, as well as other primers and probe design requirements , as the detection target sequence. According to the principle of primer probe design, use PrimerPremier5.0 software to manually design multiple sets of upstream and downstream primers and probe sequences that basically meet the requirements, and then import each set of sequences into PrimerExprssV3 software to analyze when primers and probes participate in the reaction together Whether the parameters meet the requirements, such as temperature, GC% content and other parameters, if not suitable, you need to redesign the primers or probes. Throug...

Embodiment 2

[0144] The preparation of embodiment 2 kit

[0145] The kit of this embodiment consists of the following components: sample processing solution, CPPCR reaction solution A, CPPCR reaction solution B, CP positive control and CP negative control. In order to avoid cross-contamination, the above components were prepared in different working areas, among which the sample treatment solution, CPPCR reaction solution A, CPPCR reaction solution B and CP negative control were prepared in the negative area; the CP positive control was prepared in the positive area. The preparation method of each component is as follows:

[0146] Sample treatment solution: The grades of reagents used in the preparation of the sample treatment solution are all analytically pure. After weighing the required weight, add sterilized ultrapure water to prepare a certain final concentration. The obtained sample treatment solution contains Tris-HCl (pH8.0 ) 100mmol / L; EDTA (pH8.0) 10mmol / L; NaCl1mol / L; NP-4010mg...

Embodiment 3

[0166] The preparation of embodiment 3 kits

[0167] The kit of this embodiment consists of the following components: sample processing solution, CPPCR reaction solution A, CPPCR reaction solution B, CP positive control and CP negative control. In order to avoid cross-contamination, the above components were prepared in different working areas, among which the sample treatment solution, CPPCR reaction solution A, CPPCR reaction solution B and CP negative control were prepared in the negative area; the CP positive control was prepared in the positive area. The preparation method of each component is as follows:

[0168] Sample treatment solution: The grades of reagents used in the preparation of the sample treatment solution are all analytically pure. After weighing the required weight, add sterilized ultrapure water to prepare a certain final concentration. The obtained sample treatment solution contains Tris-HCl (pH8.0 ) 100mmol / L; EDTA (pH8.0) 10mmol / L; NaCl1mmol / L; NP-4010...

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Abstract

The invention belongs to the technical field of biology and discloses a primer group and a probe for detecting chlamydia pneumoniae 98KDa MOMP genes and an application thereof. The primer group and probe consist of a forward primer with a nucleotide sequence shown in SEQ ID NO:1, a reverse primer with a nucleotide sequence shown in SEQ ID NO:2, and a probe with a nucleotide sequence shown as SEQ ID NO:3. The primer group and probe provided in the invention can be specifically combined with the chlamydia pneumoniae 98KDa MOMP genes, the detection specificity and sensitivity are obviously improved in the process of detecting the chlamydia pneumoniae 98KDa genes by adopting real-time fluorescent quantitative PCR (Polymerase Chain Reaction), and the primer group and probe can be applied to preparation of reagents for detecting the chlamydia pneumoniae 98KDa MOMP genes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a primer set and a probe for detecting the 98KDa MOMP gene of Chlamydia pneumoniae and applications thereof. Background technique [0002] Chlamydia pneumoniae (Chlamydiapneumoniae, CP) is a strictly eukaryotic intracellular parasitic prokaryotic microorganism, together with Chlamydia psittaci, Chlamydia abortus, Chlamydia guinea pigs, Chlamydia cats, and Chlamydia animals form the genus Chlamydia. The first strain of Chlamydia pneumoniae was isolated in 1965 from a chicken embryo used for the eyes of a child in Taiwan Province, tentatively named TW-183T (small T shows the prototype strain), and in 1983 from Xiya Figure 1 A strain of AR-39 was isolated from the pharynx of a college student suffering from pharyngitis. Because some of its biological characteristics were similar to Chlamydia psittaci, it was classified as the TWAR group of Chlamydia psittaci at that time. Aft...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06C12N15/11
CPCC12Q1/6851C12Q1/686C12Q2561/101C12Q2561/113C12Q2545/114
Inventor 柳辉
Owner SHENZHEN KANGMEI BIOTECH