One group of primers and probes for diagnosing penicilliosis marneffei and application thereof
A technique for penicillosis marneffei and penicillosis, which is applied in the field of primers and probes for diagnosing penicillosis marneffei, and can solve the problems of inability to meet clinical needs, lack of rapid diagnosis of penicillosis marneffei, and low positive rate.
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[0015] The present invention will be further described below.
[0016] 1. Preparation of plasmid standards containing specific gene fragments
[0017] 1.1. Genomic DNA of Penicillium marneffei was extracted as a template, and gene fragment-specific primers and probes were synthesized according to the sequence of the ITS region:
[0018] Upstream primer: 5'-CGAATCTTTGAACGCACATTG-3';
[0019] Downstream primer: 5'-TCCCCGGAGGGACCACA-3';
[0020] Probe: FAM-5'-TGCTGTGTGCCCCGGTCATCTAGC-3'-TAMRA;
[0021] Gene fragments were amplified from Genomic DNA of Penicillium marneffei by common PCR method.
[0022] 1.2. The above gene fragments were introduced into the T vector, and transformed into Escherichia coli for replication.
[0023] 1.3. Extract plasmids containing specific gene fragments and dilute them into standards with different concentration gradients.
[0024] 2. Extract DNA from human or animal serum, whole blood, bone marrow, biopsy tissue or urine.
[0025] 3. From A...
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