Fluorescence PCR method for diagnosing infection of Chlamydia trachomatis, neisseria gonorrhoeae and ureaplasma urealyticum

A technology for Ureaplasma urealyticum and Chlamydia trachomatis, which is applied in the field of fluorescent polymerase chain reaction detection, and can solve problems such as the emerging multiple fluorescent PCR technology.

Active Publication Date: 2009-12-30
CITY UNIVERSITY OF HONG KONG
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Problems solved by technology

However, due to its technical difficulty, multiplex fluoresce

Method used

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  • Fluorescence PCR method for diagnosing infection of Chlamydia trachomatis, neisseria gonorrhoeae and ureaplasma urealyticum
  • Fluorescence PCR method for diagnosing infection of Chlamydia trachomatis, neisseria gonorrhoeae and ureaplasma urealyticum
  • Fluorescence PCR method for diagnosing infection of Chlamydia trachomatis, neisseria gonorrhoeae and ureaplasma urealyticum

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Embodiment Construction

[0010] 1. Primer and probe design (see nucleotide or amino acid sequence list and Table 1) and synthesis

[0011] Both primers and probes were synthesized by professional companies. The primers were purified by PAGE, and the probes were purified by HPLC. The 5' end of the probe is labeled with a reporter group, and the 3' end is bound with a quencher group.

[0012] Table 1. Number and position of the target sequence gene library corresponding to the amplification product in the nucleotide or amino acid sequence table

[0013] genotype

Gene Bank Number (GI No.)

target sequence position

Amplified product length (bp)

CT

FM865442

2160-2649

480

NG

AM921674

60-540

481

UU

AF085729

241-720

480

[0014] 2. Preparation of Plasmid References

[0015] Use specific primers to amplify the target sequence, connect the purified PCR product to the pGEM-T vector (Promega pGEM-Tvector system ligat...

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Abstract

The invention relates to a fluorescence PCR (polymerase chain reaction) detection method for diagnosing infection of Chlamydia trachomatis (CT), neisseria gonorrhoeae (NG) and ureaplasma urealyticum (UU), which belongs to the field of nucleic acid in vitro diagnosis. The method comprises a polymerase chain reaction (PCR) system based on fluorescence PCR technology, contains a forward primer and areverse primer aiming at the CT/the NG/the UU and a fluorescent probe, and can detects DNA of three pathogens such as the CT, the NG, the UU and the like simultaneously in a reaction tub under suitable PCR condition. The method can diagnosing the infection of the CT/the NG/the UU in a clinical sample simply, conveniently and rapidly, has high sensitivity and specificity, and has important clinical value to early control and prevention of relevant genitourinary tract infections, blocking of an infection sources, and infection reduction of related pathogen.

Description

technical field [0001] The invention is a fluorescent polymerase chain reaction (PCR) detection method for simultaneously detecting chlamydia trachomatis (CT), gonorrhea (NG) and ureaplasma urealyticum (UU) infection in clinical samples. The invention belongs to the field of in vitro nucleic acid diagnosis. Background technique [0002] Sexually transmitted diseases (STDs) are a group of global infectious diseases that are mainly transmitted through sexual contact, and some STDs can be transmitted through non-sexual contact, such as sharing needles, breastfeeding or even contact. [0003] At present, the incidence of STD in my country is on the rise. The genitourinary tract infection ( Gonorrhea and non-gonococcal urethritis) have become common diseases in the departments of gynecology, dermatology and venereal diseases, and genitourinary departments in hospitals, and are the key objects of prevention and treatment in my country. Since there are mixed infections among CT, N...

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64C12R1/01C12R1/36
Inventor 杨梦甦曾志雄傅华阳
Owner CITY UNIVERSITY OF HONG KONG
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