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Primer and probe for fluorescent quantitative PCR detection of paecilomyces farinosus

A fluorescent quantitative technology for Paecilomyces farinosa, applied in the field of fluorescent quantitative PCR detection, can solve the problems of inability to carry out quantitative detection and long time consumption, and achieve the effect of improving larval survival, fast speed and strong specificity

Pending Publication Date: 2019-11-22
成都图径生物科技有限公司
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Problems solved by technology

[0004] The traditional detection methods of Paecilomyces farinosa are fungal culture and drug susceptibility test, but both have the disadvantage of being too time-consuming and cannot be quantitatively detected

Method used

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  • Primer and probe for fluorescent quantitative PCR detection of paecilomyces farinosus

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Embodiment 1 Fluorescence quantitative PCR detects Paecilomyces lividans

[0021] Get primers and probes

[0022] Obtained the Chit1 gene sequence published by Paecilomyces lividans from Genbank (accession number: DQ406584.1), and used clustalX to analyze various fungal Chit1 gene sequences published in Genbank (accession numbers: KM221003.1, KM220989.1, GU457411.1 , EF203917.2) and Paecilomyces fumigatus Chit1 gene for multiple alignment, select the more specific region as the detection target sequence, and then use Primer Express v3.0 and Primer Primer 5.0 software to design Tagman probes and specific amplification primers , the 5' end of the probe is labeled with a fluorescent group FAM, and the 3' end is labeled with a quenching group TAMRA, which was synthesized by Chengdu Qingke Biotechnology Co., Ltd.

[0023] First, the standard curve production

[0024] (1) The strains of Paecilomyces farinosa collected in the laboratory were cultured in sodium rose red nutri...

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Abstract

The invention relates to a primer and probe for fluorescent quantitative PCR detection of paecilomyces farinosus and belongs to the technical field of fluorescence quantitative PCR detection. According to the primer and the probe for fluorescent quantitative PCR detection of the paecilomyces farinosus, a nucleotide sequence of the upstream primer is shown as SEQ ID No.1; a nucleotide sequence of the downstream primer is shown as SEQ ID No.2; a nucleotide sequence of the probe is shown in SEQ ID No.3, wherein a fluorescent reporter group is bound to the 5' terminal of the nucleotide sequence ofthe probe and a fluorescent quenching group is bound to the 3' terminal of the nucleotide sequence of the probe. The invention further provides a detection method of the paecilomyces farinosus. By applying the primer and the probe, the rapid quantitative detection of the paecilomyces farinosus can be realized, the pollution of the paecilomyces farinosus can be reduced in the artificial cultivation process of cordyceps sinensis, the survival rate of swift moth larvae is improved, and a powerful guarantee is provided for artificial large-scale production of cordyceps sinensis.

Description

technical field [0001] The invention relates to primers and probes for detecting Paecilomyces lividans by fluorescent quantitative PCR, and belongs to the technical field of fluorescent quantitative PCR detection. Background technique [0002] Isaria farinosa, also known as Cordyceps Cordyceps, is a soil-dwelling fungus. According to research, it can infect almost all insects and is a common entomopathogenic fungus. Because Paecilomyces trichoderma is widely distributed in the world and has a wide host range, it has been used as a biological factor to control agricultural pests. [0003] Although Paecilomyces trichoderma is beneficial in biological control, it is a highly lethal pathogen of the host insect bat moth in the artificial cultivation process of the precious traditional Chinese medicine Cordyceps sinensis, which directly affects the survival rate of larvae, and has been found in many years of practice. The occurrence of diseases is increasing, which has caused pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6851C12Q2531/113C12Q2563/107C12Q2545/114
Inventor 林冬梅杨涛杨妙婷魏巍
Owner 成都图径生物科技有限公司
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