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.

Active Publication Date: 2014-01-15
GUANGXI MEDICAL UNIVERSITY
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Problems solved by technology

At present, the diagnosis of Penicillium marneffei still relies on traditional fungal culture and morphological identification. The culture takes 1-4 weeks, which takes a long time and the positive rate is relatively low, which is far from meeting the clinical needs.
There is currently no preparation for the rapid diagnosis of Penicillium marneffei

Method used

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  • One group of primers and probes for diagnosing penicilliosis marneffei and application thereof
  • One group of primers and probes for diagnosing penicilliosis marneffei and application thereof
  • One group of primers and probes for diagnosing penicilliosis marneffei and application thereof

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Embodiment Construction

[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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Abstract

The invention discloses one group of primers and probes for diagnosing penicilliosis marneffei. The sequence of the upstream primer is SEQ NO.1: 5'-CGAATCTTTGAACGCACATTG-3'; the sequence of the downstream primer is SEQ NO.2: 5'-TCCCCGGAGGGACCACA-3'; the sequence of the probes is SEQ NO.3: FAM-5'-TGCTGTGTGCCCCGGTCATCTAGC-3'-TAMRA. The primers and the probes can specifically detect genomic DNA of the penicilliosis marneffei, and are applied to detection of DNA capacity of the penicilliosis marneffei in serum, whole blood, marrow or urine of human or animals, necessary conditions for research and development of reagent for diagnosing and estimating the penicilliosis marneffei are created, and the important theoretic and practical significance is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a set of primers and probes for diagnosing penicillium marneffei. technical background [0002] Penicillium marneffei occurs in Southeast Asia and is one of the most common opportunistic infections among AIDS patients in southern my country. It is an important factor leading to the death of local AIDS patients, with a mortality rate of 80%. The onset of Penicillium marneffei is insidious, with complex clinical manifestations and no obvious features. However, if it can be diagnosed early and treated with antifungal drugs in time, good curative effect can usually be obtained. At present, the diagnosis of Penicillium marneffei still relies on traditional fungal culture and morphological identification. The culture takes 1-4 weeks, which takes a long time and the positive rate is relatively low, which is far from meeting the clinical needs. There is currently no preparation for the rapi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06C12N15/11G01N21/64
CPCC12Q1/04C12Q1/686C12Q2563/107C12Q2537/16
Inventor 曹存巍莫冬冬梁伶林有坤梁浩
Owner GUANGXI MEDICAL UNIVERSITY
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