Fluorescent quantitative PCR detection kit for chlamydia trachomatis

A technology for Chlamydia trachomatis and detection kit, which is applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of unsuitable clinical diagnosis and diagnosis basis, relying on subjective judgment, poor detection accuracy, etc., and achieves fast operation. Clear, reduced risk of contamination, easy to operate

Inactive Publication Date: 2015-11-04
兰州安康伯乐生物技术有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

The culture method is accurate and reliable, but it often takes 2-3 days, the culture period is long, and the operation is cumbersome. Due to the influence of sample collection methods and transportation methods, it often leads to shortcomings such as low positive rate, which can no longer meet the pace of rapid diagnosis and treatment of modern medicine; direct The smear method is simple to operate, but the detection accuracy is poor and depends on subjective judgment; although immunological methods such as colloidal gold technology and enzyme-linked immunosorbent assay have the advantages of simplicity and speed, they have poor specificity, low sensitivity, and high false positive rate. Disadvantages: It is not suitable for clinical diagnosis and confirmation, and can only be used as a tool for initial diagnosis

Method used

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  • Fluorescent quantitative PCR detection kit for chlamydia trachomatis
  • Fluorescent quantitative PCR detection kit for chlamydia trachomatis
  • Fluorescent quantitative PCR detection kit for chlamydia trachomatis

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Embodiment 1

[0033] The preparation of embodiment 1 kit

[0034] 1、 Design and synthesis of kit-specific primers

[0035] According to the CT sequence published online (GenBank: CP006677.1), the Chlamydia trachomatis PCR primers were designed for the conserved Chlamydia trachomatis gene trpB fragment using the biology software Primer Premier 5 software and Oligo 7 software, and the software Mega4 and NCBI online Blast were used for the design Sequence alignment ensures the specificity and rationality of each pair of primers. The primers were synthesized by PAGE Bioengineering (Dalian) Co., Ltd. and purified by PAGE. The nucleotide sequences of the kit-specific primers are as follows:

[0036] Forward primer: 5'-GCATGAGTCAGGACGAGTT-3',

[0037] Reverse primer: 5'-CGAAACTAAATGTGCGAGAGC-3',

[0038] Fluorescent probe: 5'-FAM-TTAGCCACCGATGAAGACCCGTTAC-TAMRA-3',

[0039] 2. Design and preparation of CT plasmid positive quality control products

[0040] A pair of amplification primers ar...

Embodiment 2

[0056] The usage method of embodiment 2 kit

[0057] 1. Sample requirements

[0058] a. Sample collection: A special swab sterilized by autoclaving is inserted into the male urethral orifice or female cervix 1-2 cm, rotated once, take it out after staying for about 10 seconds, put the cotton swab head into 1 ml sterile In the cryopreservation tube of the sample of normal saline, cut the swab from the neck, immerse the swab head in the saline, and rinse repeatedly to obtain the specimen rinsing solution.

[0059] b. Storage: The specimen rinse solution should be stored below -20°C, and the samples that cannot be detected within three months should be stored below -70°C; the specimens should be stored separately in a special cabinet or warehouse.

[0060] c. Transportation: Use curlers with ice packs or foam boxes with ice packs for transportation.

[0061] 2. Sample preparation is the sample processing area

[0062] Take 500 μl of sample washing solution, centrifuge at 10,00...

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Abstract

The invention provides a fluorescent quantitative PCR detection kit for chlamydia trachomatis in clinic samples, used for assisted diagnosis of chlamydia trachomatis. The kit comprises a PCR solution, a DNA polymerase solution, positive quality control, weakly positive quality control, negative quality control, positive quantitative reference, lysate and protease K, wherein the PCR solution contains forward and reverse primers and the fluorescence probe are specific primers and probe designed for the specific sequence of chlamydia trachomatis and are capable of amplifying a target DNA sequence specifically so as to conveniently and quickly detect chlamydia trachomatis infection in clinic samples. The kit has the characteristics of high specificity and high sensitivity.

Description

technical field [0001] The invention belongs to the field of in vitro nucleic acid detection, and the invention provides a method for detecting Chlamydia trachomatis (Chlamydia trachomatis) in clinical samples Chlamydia trachomatis , referred to as CT) fluorescent quantitative PCR kit for early clinical diagnosis of Chlamydia trachomatis. Background technique [0002] Chlamydia trachomatis ( Chlamydia trachomatis , CT) is a class of Chlamydia, between viruses and Rickettsia, can pass through bacteria filters, prokaryotic microorganisms with obligate intracellular parasitism and unique life cycle. Chlamydia has affinity for epithelial cells on the mucosa, especially columnar epithelial cells, so it is easy to cause infection of the ocular conjunctiva and genitourinary system mucosa. [0003] Chlamydia trachomatis infection ranks third among sexually transmitted diseases (STDs) in my country, and its incidence is increasing year by year. At present, Chlamydia trachomatis in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06
Inventor 杨金波王黎明刘志新杜宇平刘音希
Owner 兰州安康伯乐生物技术有限公司
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