Pathogen nucleic acid-drug resistant gene detection kit and application thereof
A gene detection and drug resistance technology, applied in the field of medicine and biology, can solve the problems of no mycoplasma resistance detection reagents, multiple target genes or pathogen detection, etc., and achieve rapid and objective detection results, strong repeatability and sensitivity high effect
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Embodiment 1
[0065] The kit includes the following components:
[0066] PCR reaction solution, enzyme mixture, Chlamydia trachomatis / Ureaplasma urealyticum / Mycoplasma hominis / drug resistance gene multiple reaction solution, positive control 1, negative control;
[0067] The PCR reaction solution includes 10× buffer, 25mM MgCl2, 10mM dUTP and 10mM dNTPs;
[0068] The enzyme mixture includes Taq enzyme and UNG enzyme, the Taq enzyme is a hot start Taq enzyme, and the UNG enzyme is uracil-N-glycosylase.
[0069] Chlamydia trachomatis / Ureaplasma urealyticum / Mycoplasma hominis / drug resistance gene multiplex reaction solution upstream and downstream primers, and the ratio of the probe is: 6:4:2; the preferred upstream primer is 600nM, the downstream primer is 400nM, the probe is 200nM.
[0070] The nucleotide sequences of primers and probes of Mycoplasma hominis, Ureaplasma urealyticum, Chlamydia trachomatis and Ureaplasma urealyticum tetracycline resistance gene used in this example can be ...
Embodiment 2
[0110] The kit includes the following components:
[0111] PCR reaction solution, enzyme mixture, Chlamydia trachomatis / Ureaplasma urealyticum / Mycoplasma hominis multiplex reaction solution, drug resistance gene reaction solution, positive control 1, positive control 2, negative control;
[0112] The PCR reaction solution includes 10× buffer, 25mM MgCl2, 10mM dUTP and 10mM dNTPs;
[0113] The enzyme mixture includes Taq enzyme and UNG enzyme, the Taq enzyme is a hot start Taq enzyme, and the UNG enzyme is uracil-N-glycosylase.
[0114] Chlamydia trachomatis / Ureaplasma urealyticum / Mycoplasma hominis multiplex reaction solution has a ratio of upstream and downstream primers and probes of 6:4:2; the preferred upstream primer is 600nM, the downstream primer is 400nM, and the probe is 200nM.
[0115] The ratio of the upstream and downstream primers and probes of the drug resistance gene reaction solution is: 6:4:2; the preferred upstream primer is 600nM, the downstream primer is...
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