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Triple nucleic acid test kit

A detection kit and triple nucleic acid technology, applied in the field of nucleic acid detection of fluorescent quantitative PCR

Active Publication Date: 2017-01-11
BEIHAI SINLON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] By comparison, the above-mentioned patented technical schemes disclosed above are substantially different from the quantitative detection scheme of the present application which mixes three common pathogenic bacteria in one system and can distinguish the types of bacteria

Method used

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example

[0051] Example: Preparation of kits

[0052] (1) Primer and probe design and synthesis

[0053] The specific primer sequences are highly conserved and specific sequences for the ITS of Aspergillus, Cryptococcus neoformans, and Candida albicans, and the labeled reporter groups of the three specific probes are FAM, VIC, and CY respectively. -5, the quenching group is MGB, and the data are all derived from the ribosomal ITS sequence in GenBank, and the primers and probes are synthesized by life technologies.

[0054] (2) Preparation of positive quantitative reference substance and negative reference substance

[0055] Using the extracted Aspergillus genome, Cryptococcus neoformans genome, and Candida albicans genome as templates, the positive reference products of Aspergillus were amplified using Aspergillus primers, dNTPs, dUTP, 10*taq buffer, and Taq enzyme as reaction systems. Cryptococcus primers, dNTPs, dUTP, 10*taq buffer, and Taq enzyme were used as the reaction system t...

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Abstract

The invention relates to a triple nucleic acid test kit. The kit comprises a thallus lysate, a positive quantitative reaction liquid, a negative control reaction liquid and a thallus test reaction liquid, wherein the thallus test reaction liquid contains specific primers, specific probes and doping fluorescent dye for aspergillus, Cryptococcus neoformans and candida albicans, the three specific oprimers are designed according to areas of ITS sequences of aspergillus, Cryptococcus neoformans and candida albicans, FAM, VIC and CY-5 are selected for mark report groups of the three specific probes, MGB serves as a quenching group, the excitation wavelength of the FAM group is 492 nm, and the emission wavelength is 517 nm; the excitation wavelength of the VIC group is 557 nm and the emission wavelength is 574 nm; the excitation wavelength of the CY-5 group is 646 nm and the emission wavelength is 664 nm. The triple nucleic acid test kit for aspergillus, Cryptococcus neoformans and candida albicans can assist in diagnosing infection caused by aspergillus, Cryptococcus neoformans and candida albicans.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection by fluorescence quantitative PCR, in particular to a triple nucleic acid detection kit. Background technique [0002] At present, deep fungal infections are mostly caused by opportunistic pathogenic fungi, which are more common in immunocompromised populations and have a high mortality rate. Among them, invasive fungal infections caused by Candida accounted for 70% to 90% of all cases, among which Candida albicans accounted for 70 to 80% of Candida infections; Aspergillus infection accounted for 10% to 20% of fungal infections; and Cryptococcus Among them is mainly Cryptococcus neoformans, which is a common pathogenic fungus co-infected by AIDS patients, causing cryptococcal meningitis in AIDS patients, and the cure rate is extremely low. [0003] At present, there are five types of antifungal drugs commonly used in clinical practice, polyenes, azoles, echinocandins, flucytosines, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/66C12R1/645C12R1/725
CPCC12Q1/6895
Inventor 李可可韩钊任卫新李智鑫王楠李松强苑庆华辛鹤林
Owner BEIHAI SINLON BIOTECH CO LTD
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