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Double qPCR method for rapidly detecting chlamydia trachomatis, primer and probe

A Chlamydia trachomatis, rapid technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of lack, missed diagnosis and misdiagnosis of Chlamydia trachomatis infection, and achieve simple patents, improved diagnostic capabilities, and high sensitivity Effect

Pending Publication Date: 2019-05-14
深圳市宝安区沙井人民医院 +1
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Problems solved by technology

However, the qPCR method with high sensitivity and high specificity that can be used for the detection of Chlamydia trachomatis is still lacking, which leads to the missed diagnosis and misdiagnosis of Chlamydia trachomatis infection

Method used

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  • Double qPCR method for rapidly detecting chlamydia trachomatis, primer and probe
  • Double qPCR method for rapidly detecting chlamydia trachomatis, primer and probe
  • Double qPCR method for rapidly detecting chlamydia trachomatis, primer and probe

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Embodiment Construction

[0045] The present invention will be further described below in conjunction with accompanying drawing and specific embodiment:

[0046] Such as figure 1 , figure 2 , image 3As shown, a double qPCR method for rapid detection of Chlamydia trachomatis comprises the following steps:

[0047] (1) Preliminary preparation procedures:

[0048] Research samples: Randomly collect several urogenital tract samples from patients with sexually transmitted diseases, including several male urethral swabs, female endocervical swabs and vaginal swabs, for method evaluation.

[0049] Instruments and reagents:

[0050] ①Nucleic acid extractor (ExPure20) and its supporting magnetic bead extraction and purification reagents;

[0051] ② Fluorescent quantitative PCR instrument (LightCycler480); multiplex fluorescent quantitative PCR system (Platinum TM Multiplex PCR Master Mix);

[0052] ③PCR product sequencing;

[0053] (2) Nucleic acid extraction and purification:

[0054] ①Original samp...

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Abstract

The invention provides a double qPCR method for rapidly detecting chlamydia trachomatis, a primer and a probe, wherein the method comprises the following steps: (1) preliminary preparation procedure;(2) extraction and purification of nucleic acid; (3) primer / probe design, synthesis and labeling; (4) qPCR amplification and result judgment; (5) testing the detection lower limit and the amplification linear range; (6) calculation of diagnostic sensitivity and specificity; (7) statistical methods; the double qPCR method for rapidly detecting chlamydia trachomatis has the advantages that the detection lower limit is as low as 2copies / PCR, the amplification linear range reaches 1.0*102-1.0*108 copies / mL, the diagnostic sensitivity and specificity for the chlamydia trachomatis are 100.0%(134 / 134) and 99.3%(1142 / 1150) respectively, and the report result is less than or equal to 2.0 hours from sample treatment; the method has the advantages of simplicity, convenience, rapidness, high sensitivity and good specificity, not only can improve the diagnosis capability of chlamydia trachomatis infection, but also can achieve rapid detection and gain time for early accurate treatment.

Description

【Technical field】 [0001] The invention relates to a double qPCR method for chlamydia trachomatis, in particular to a double qPCR method for rapid detection of chlamydia trachomatis and primers and probes. 【Background technique】 [0002] Chlamydia trachomatis (Chlamydia trachomatis, CT) is an obligate intracellular parasitic pathogen, and different serotypes can cause different diseases and complications, including reproductive tract infection (serotype D-K), trachoma (serotype A-C) and venereal lymphatic Granulomas (serotypes L1-L3) [1-3]. As the most common sexually transmitted pathogen, Chlamydia trachomatis can cause severe pelvic inflammatory disease, ectopic pregnancy, and tubal infertility [4], bringing enormous pressure and medical consumption to families and society. Every year, there are about 92 million new cases of Chlamydia trachomatis infection in the genitourinary tract worldwide, and about 50% of male infected people and 70% of female infected people do not s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/01
Inventor 刘和录马彩凤杜纪坤窦宇红何维娜袁晓雪李玉霞赵丽君刘平
Owner 深圳市宝安区沙井人民医院
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