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.
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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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