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PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method for chlamydia trachomatis

A Chlamydia trachomatis and detection kit technology, which is applied in the direction of microbial measurement/inspection, fluorescence/phosphorescence, biochemical equipment and methods, etc., can solve the problems of cumbersome operation and operational pollution, and achieve quantitative accuracy, high accuracy, and sensitivity and the effect of precision improvement

Active Publication Date: 2013-09-18
泰普生物科学(中国)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Huamei Biotech uses traditional PCR technology to amplify the CT target gene first, then combines the amplified product with the ELISA method to hybridize with the specific probe fixed on the microplate plate, and then uses the ELISA method to carry out chromogenic titration for CT- DNA quantification, the advantage of this method is that no additional fluorescent PCR instrument is required, but this method is cumbersome to operate and easily causes operational pollution

Method used

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  • PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method for chlamydia trachomatis
  • PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method for chlamydia trachomatis
  • PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method for chlamydia trachomatis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Composition and configuration of embodiment 1 kit

[0042] 1. Main raw material sources and preparation methods

[0043] The CT primers and probe sequences of this kit are as follows:

[0044] Upstream primer: 5′-CTGTACATTAGGAGCCACCA-3′;

[0045] Downstream primer: 5′-CAACAGATTGATCAAAGTC-3′;

[0046] Fluorescent probes:

[0047] 5'-FAM-GATATCTTAAAGGAAATTCAGCATCTTTCAA-TAMRA-3'.

[0048] The internal reference (Suc) primer upstream primer sequence of this kit: SEQ ID NO.4, downstream primer sequence: SEQ ID NO.5 and probe sequence: SEQ ID NO.6, specifically as follows:

[0049] Upstream primer: 5′-TCATCGTCGCTGGAGCTGGTT-3′;

[0050] Downstream primer: 5′-CGGCGGTTTGTCAAGCTGAT-3′;

[0051] Fluorescent probes:

[0052] 5'-HEX-CTTCTTATAGTCACTGCACTAAACTGGAT-TAMRA-3'.

[0053] The kit also includes DNA polymerase, strong positive quality control substance, weak positive quality control substance, negative quality control substance, positive quantitative reference substan...

Embodiment 2

[0094] The use of embodiment 2 kit

[0095] a. Reagent preparation (reagent preparation area)

[0096] 1. Take out the DNA extraction solution and set aside.

[0097] 2. After determining the number of reaction tubes n (number of samples + negative + positive quality control + weak positive quality control), take out the CT-PCR reaction solution and absorb n×44μl CT-PCR reaction solution, n×1μl DNA polymerization For the enzyme system, add to a centrifuge tube and vortex to mix well. After brief centrifugation, divide into n PCR reaction tubes, 45 μl per tube. Cover the tube cap and transfer to the sample loading area, and store in a 4°C refrigerator in the dark for future use.

[0098] 3. Transfer the quality control substance and quantitative reference substance to the sample processing area, and put them in a 4°C refrigerator for later use.

[0099] b. Sample processing (sample processing area)

[0100] sample requirements

[0101] 1. Applicable sample types: reproducti...

Embodiment 3

[0145] Embodiment 3 This kit performance evaluation

[0146] The DNA concentration of the positive quantitative reference product is P1: 1.0×10 7 copies / ml, P2: 1.0×10 6 copies / ml, P3: 1.0×10 5 copies / ml, P4: 1.0×10 4 copies / ml.

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Abstract

The invention aims at providing a kit which is suitable for rapid test of chlamydia trachomatis in clinical samples and can be used for auxiliary diagnosis and efficacy monitoring of chlamydia trachomatis infection. The technical scheme of the invention is as follows: an PCR (polymerase chain reaction) fluorescence quantitative rapid test kit of the chlamydia trachomatis is provided and comprises a PCR reaction solution, wherein the PCR reaction solution contains primers and a fluorescence probe; and the primers comprise an upstream primer and a downstream primer, and the kit further comprises a DNA (deoxyribonucleic acid) polymerase, a strong positive quality control product, a weak positive quality control product, a negative quality control product, a positive quantitative reference product and a DNA extraction solution. According to the CT (chlamydia trachomatis) fluorescence PCR quantitative test kit and method provided by the invention, a Taqman core technology platform and an arabidopsis internal reference system are utilized, thus the test sensitivity is higher. Furthermore, the accuracy, specificity, repeatability, stability, sensitivity and precision are improved compared with those of the existing product.

Description

technical field [0001] The invention relates to a disease pathogen gene detection technology, in particular to a PCR fluorescence quantitative rapid detection kit for Chlamydia trachomatis and a method thereof. Background technique [0002] Chlamydia trachomatis is a kind of prokaryotic cell-type microorganisms that can pass through bacteria filters, strictly eukaryotic intracellular parasitism, and have a unique development cycle. It can cause trachoma, inclusion body envelope inflammation, genitourinary tract infections, and lymphogranuloma venereum. After injection of Chlamydia trachomatis vaccine, local specific immunity can be induced, but its protective effect can only last for 1-2 years. According to "Berger's System Handbook" (1984), Chlamydia trachomatis is divided into three biotypes, namely, biovar mouse, biovar trachoma and biovarlymphogranulomavenereum. , LGV), the latter two are related to human diseases. Using indirect micro-immunofluorescence test, the tra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06G01N21/64
Inventor 何东华蔡晓沂
Owner 泰普生物科学(中国)有限公司
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