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Quality control substance for detecting chlamydi trachomatis

A technology for Chlamydia trachomatis and quality control substances, which is applied in the fields of clinical laboratory science and biology, can solve the problem of restricting the quality assurance system of pathogen detection, being unsuitable for large-scale clinical detection quality control work, increasing operational complexity and manufacturing costs. The risk of biological infection and other problems, to achieve the effect of practicality, wide applicability, and reduction of the generation of enzyme cleavage sites.

Inactive Publication Date: 2009-02-11
BEIJING HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(3) Vaginal specimens are not suitable for this examination
No matter which of the above methods requires special treatment of samples, increases operational complexity and production costs, and has a certain risk of biological infection, it is not suitable for large-scale clinical testing quality control work, thus virtually limiting the detection of pathogens. Improvement of quality assurance system

Method used

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  • Quality control substance for detecting chlamydi trachomatis
  • Quality control substance for detecting chlamydi trachomatis
  • Quality control substance for detecting chlamydi trachomatis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Construction of recombinant vectors containing CT cryptic plasmid fragments

[0054] 1. Experimental materials:

[0055] 1. Source of specimens: Chlamydia trachomatis cell cultures were preserved in our laboratory, and cervical epithelial cells (HTB-Siha cells) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0056] 2. Main reagents:

[0057] Bacterial DNA Extraction Kit Tiangen Biochemical China

[0058] Spin Column Type Ordinary Gel DNA Recovery Kit Tiangen Biochemical China

[0059] Taq polymerase Promega Inc. USA

[0060] DNA Electrophoresis Marker: DL2000 Takara Japan

[0061] DNA Electrophoresis Marker: DL15000 Takara Japan

[0062] TA Cloning Kit (T-Easy Vector) Promega, Inc. USA

[0063] Plasmid (miniature) extraction kit Promega USA

[0064] Restriction enzyme Sal I New England Biology USA

[0065] Restriction enzyme Not I New England Biology USA

[0066] T4DNA Ligase Promega, Inc., USA

[0067] Strain: DH5 α...

Embodiment 2

[0374] Example 2: Stability study of quality control substances and external quality evaluation of CT PCR detection applied in clinical laboratories

[0375] 1. Experimental materials

[0376] 1. Specimen source: Chlamydia trachomatis PCR detection quality control material constructed in Example 1

[0377] 2. Main reagents:

[0378] Cell culture medium DMEM (high glucose) Hyclone Corporation USA

[0379] Fluorescent quantitative RT-PCR detection reagent for Chlamydia trachomatis Guangzhou Daan Company China

[0380] Chlamydia trachomatis Fluorescent Quantitative RT-PCR Detection Reagent Shenzhen Piji Company China

[0381] 3. Main instruments:

[0382] Real-time fluorescence PCR instrument (7500Real Time PCR System) American biological application system United States

[0383] Real-time fluorescent PCR instrument (Lightcycler) Roche Company USA

[0384] Real-time fluorescent PCR instrument (Line-gene PCR instrument) Hangzhou Bioer Technology Co., Ltd. China

[0385] Des...

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Abstract

The invention discloses a quality control substance used for testing chlamydia trachomatis, belonging to the field of clinical ecsomatics and biotechnology. The quality control substance used for testing chlamydia trachomatis contains nucleotide sequence or complementary sequence thereof which is shown by sequence tables of SEQ ID No.1 to SEQ ID No.4. The quality control substance has the advantages that (1) by adopting overlapping PCR and the method of double enzyme digestion, a simple and effective method for connecting and constructing segments with a plurality of intervals into the same carrier is set up, and the probability of lower efficiency in long distance PCR is avoided by directed cloning, so that the two methods can be combined for use to make up for the deficiencies of each other. (2) Clinical specimen which can be simulated and has no biological infection fatalness is successfully obtained, and the quality control substance for testing polymerase chain reaction of the chlamydia trachomatis can be produced in large quantity, is stored stably and has better applicability.

Description

technical field [0001] The invention relates to a quality control substance for detecting chlamydia trachomatis, belonging to the fields of clinical laboratory science and biotechnology. Background technique [0002] Chlamydia trachomatis (CT) is one of the pathogens of important sexually transmitted diseases. It not only causes vaginal and urinary tract infections; ascending reproductive tract infections may also involve the endometrium, fallopian tubes and adjacent pelvic structures, and can also cause Pelvic inflammatory disease, fallopian tube damage until infertility. In addition, this pathogen is also associated with human papillomavirus (HPV) and human immunodeficiency virus (HIV) infections. [0003] Chlamydia is a Gram-negative pathogen. It is an obligate intracellular parasitic microorganism. It has no ability to synthesize the high-energy compound ATP and must be provided by the host cell, so it is an energy parasite. Chlamydia is a prokaryotic microorganism tha...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12N15/85
Inventor 李金明霍虹王露楠张括张瑞
Owner BEIJING HOSPITAL
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