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Kit for detecting sexually transmitted diseases

A technology for transmitting diseases and kits, which is applied in the field of detection of sexually transmitted diseases, can solve problems such as the absence of a silver staining kit for gonococcus, and achieve the effects of low requirements, low cost and simple steps.

Inactive Publication Date: 2011-06-01
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0006] There is no silver staining kit for detecting Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Ureaplasma urealyticum (UU) at home and abroad
At the same time, there is no method to simultaneously detect the three pathogenic DNAs of Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Ureaplasma urealyticum (UU) using multiplex PCR technology and use silver staining method for separation and detection

Method used

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  • Kit for detecting sexually transmitted diseases
  • Kit for detecting sexually transmitted diseases
  • Kit for detecting sexually transmitted diseases

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Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1 Primer design and synthesis of the present invention

[0047] 根据淋球菌(NG),沙眼衣原体(CT)和解脲支原体(UU)的特征基因,即:淋球菌Pjd1质粒基因:674bp-844bp产物长度171bp,序列如SEQ ID NO:7所示(即序列cctatcgggctgttatctgattatggttattttgacatagttgtatcatcttaaaaacaagcatacaatctgcaaatcttagacaaagcaaaacccccgccaaacgccaatctgcacgggggtttcgagatacaacatgagccaattatacacccaacccgacctcttctt );沙眼衣原体Plgv440隐蔽性质粒基因:63bp-269bp产物长度207bp,序列如SEQ ID NO:8所示(即序列cttccccagaacaataagaacacacactttgtctcgatgagagacaggaaatacgcatgatttcctca tcttttaatcctatttgctttaaatgaatcaaagcgcttgcacgaagtactctgggagtaatttttattttcatagcactatggaactctgcaagcctaaaattatgcgcaacctgacttgttgttacaggaatcccta);解脲支原体16S rRNA编码区V5和V7区域:432bp-732bp产物长度 313bp(即序列taaatgtcggctcgaacgagtcggtgttgtagctaacg cattaaatgatgtgcctgggtagtacattc gcaagaatgaaactcaaacggaattgacggggacccgcacaagtggtggagcatgttgcttaatttgacaatacacgtagaaccttacctaggtttgacatctattgcgacgctatagaaatatagttgaggttaacaatatgacaggtggtgcatggttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccctttcgttagttgcttttctagcgatactgc),序列如SEQ ID N...

Embodiment 2

[0055] The specificity experiment of embodiment two primers of the present invention

[0056] 1. Source of specimens: three standard strains of Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Ureaplasma urealyticum (UU) were provided by the Microbiology Laboratory of West China Hospital of Sichuan University, and temporarily stored at -20 degrees Celsius after making suspensions;

[0057] 2. Materials and methods:

[0058] Reagents: (1) DNA Lysis Solution:

[0059] Also known as pre-reaction solution, it contains: 50mmol / L NaOH, 10mmol / L Tris-HCL, pH 8.0, volume fraction 1% Triton X-100, volume fraction 1% NP-40, 0.5mmol / L EDTA pH 8.0.

[0060] (2) Amplification reaction tube:

[0061] In the present invention, the amplification reaction tube is a 75ul reaction system, which specifically includes: Reaction buffer (10*) 7.5ul; DNTP (10mmol): 1.5ul; Taq enzyme [TaKaRa Bio-technology (Dalian) CO.Ltd] ( 5U / ul): 1.5ul; Mg 2+ (25mmol / ul): 4.5ul; Primer (20P): Neisseri...

Embodiment 3

[0077] Embodiment 3 Preparation of detection kit of the present invention

[0078] composition:

[0079] 1. Primers:

[0080] SEQ ID NO: 15'CCTATCGGGCTGTTATCTGATTATGG 3'

[0081] SEQ ID NO: 25'AAGAAGAGGTCGGGTTGGGTGTAT 3'

[0082] SEQ ID NO: 35'CTTCCCCAGAACAATAAGAACACA 3'

[0083] SEQ ID NO: 45'TAGGGATTCCTGTAACAACAAGTCA 3'

[0084] SEQ ID NO: 55'TAAATGTCGGCTCGAACGAG 3'

[0085] SEQ ID NO: 65'GCAGTATCGCTAGAAAAGCAAC 3'

[0086] 2. DNA lysate:

[0087] Also known as pre-reaction solution, it contains: 50mmol / L NaOH, 10mmol / L Tris-HCL, pH 8.0, volume fraction 1% Triton X-100, volume fraction 1% NP-40, 0.5mmol / L EDTA pH 8.0.

[0088] 3. Amplification reaction tube:

[0089] In the present invention, the amplification reaction tube is a 75ul reaction system, which specifically includes: Reaction buffer (10*) 7.5ul; DNTP (10mmol): 1.5ul; Taq enzyme [TaKaRa Bio-technology (Dalian) CO.Ltd] ( 5U / ul): 1.5ul; Mg 2+ (25mmol / ul): 4.5ul; Primer (20P): Neisseria gonorrhoeae NG, Sense...

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Abstract

The invention discloses a kit for detecting sexually transmitted diseases. The kit comprises 1 to 3 pairs of primers which can specifically amplify genes of pathogens such as gonococcus, chlamydia trachomatis and / or ureaplasma urealyticum respectively; and the nucleotide sequences of the primers are shown as SEQ ID NO:1-6. The kit has high specificity and high sensitivity, can be used for detecting a single pathogen and detecting various pathogens at the same time, is convenient to use, provides exact results and has an extremely good clinical application prospect.

Description

technical field [0001] The invention relates to a detection technology for sexually transmitted diseases, in particular to a detection method for detecting pathogens in human genital secretions, a kit and corresponding primers. Background technique [0002] Sexually transmitted disease (STD) is not only one of the oldest human diseases, but also the most widespread infectious disease in the world. STD is a kind of disease that is mainly transmitted through sexual contact between infected and uninfected people. Its concept has been expanded from the past classic diseases caused by 5 pathogens to more than 20 pathogens closely related to sexual contact. caused by infectious diseases. In recent years, the incidence of STD has shown an upward trend. From 1980 to 2000, there were only 48 cases of STD infection reported nationwide. STD spread rapidly in my country. STD can cause male and female genitourinary tract infection, cause infertility and even canceration. The current p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 应斌武刘嘉铭邹秀和黄进王兰兰
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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