Primer probe group and kit for combined detection of mycoplasma pneumoniae and chlamydia pneumoniae based on fluorescence RMA method
A technology of mycoplasma pneumoniae and chlamydia pneumoniae, applied in the direction of microorganism-based methods, DNA/RNA fragments, microorganisms, etc., can solve the problems of easy pollution, poor sensitivity, multiple false positive results, etc., to avoid false positives and environmental pollution, Reduce reaction time and temperature, avoid cross-reaction effects
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Embodiment 1
[0045] 1. Preparation of positive standard plasmid
[0046] The RNA of MP and Cpn was extracted as a template, and the adhesion protein P1 gene of MP and the outer membrane protein A gene of Cpn were amplified by PCR. The PCR amplification products were subjected to 1% agarose gel electrophoresis, recovered by tapping gel, and cloned into pMD18 -T vector, transformed into Escherichia coli competent cells, blue-white screening, picking white colonies, and colony PCR verification. Send the positive recombinant bacteria to the company for sequencing, culture the correctly sequenced recombinant bacteria overnight, extract plasmid DNA, and obtain positive plasmids (P-MP, P-Cpn).
[0047] 2. Design of fluorescent RMA primers and probes
[0048] Fluorescent RMA primers and probes were designed for the adhesion protein P1 gene of MP and the outer membrane protein A gene of Cpn, as shown in Table 2:
[0049] Table 2 Primer and Probe Sequences
[0050]
[0051] Note: The fluoresce...
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Description
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Application Information
- IPC
- C12Q1/689; C12Q1/6844; C12Q1/04; C12N15/11; C12R1/35; C12R1/01
- CPC
- C12Q1/689; C12Q1/6844
- Inventors
- 刘建生; 包建民