Triple PCR detection method for rapidly distinguishing non-LP (legionella pneumophila), LP and LP I
A detection method and technology for Legionella, which is applied in the field of multiplex PCR detection of Legionella, can solve the problems of uncertainty, multiple cross-reactivity, and doubtful Legionella colonies cannot be finally identified, and achieves low cost, accurate and reliable results, The effect of the simple method
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[0017] Example 1 A triple PCR detection method for rapidly distinguishing non-pneumophilic, pneumophilic and pneumophilic type 1 Legionella, the steps are:
[0018] 1. Prepare a PCR template, pick a suspicious single colony to be detected and put it in 0.5mL sterilized nuclease-free deionized water to prepare a bacterial suspension, boil it at 100°C for 15min, and cool it for later use.
[0019] 2. Primer sequence:
[0020] 16S rRNA gene: upstream primer 5'-AAGATTAGCC TGCGTCCGAT-3', downstream primer 5'-GTCAACTTAT CGCGTTTGCT-3', the expected length of the amplified fragment is 654 bp;
[0021] danJ gene: upstream primer 5'-AGGTGGTTTT GGCGGATTTG G-3', downstream primer 5'-TGAATTCTGA CTTGCCCCAT G-3', the expected length of the amplified fragment is 285 bp;
[0022] ORF9 gene: upstream primer 5'-CAGGATTACC GCTCATTATT G-3', downstream primer 5'-GTAATTCCCA GCCATTTACC AGATC-3', the expected length of the amplified fragment is 561 bp.
[0023] 3. PCR reaction system: 1.6 μL d...
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