Multiplex quantitative PCR (Polymerase Chain Reaction) primer, probe, kit and detection method for corynebacterium bacteria and Klebsiella corynebacterium

A technology of multiple fluorescent quantification and coryneform bacteria, which is applied in the fields of molecular biology and medicine, can solve the problems of unsatisfactory clinical diagnosis and treatment, low positive rate, and poor accuracy, and achieve the goal of short detection time, favorable price, and high accuracy Effect

Active Publication Date: 2017-10-10
GUANGDONG WOMEN & CHILDREN HOSPITAL
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Problems solved by technology

Since Corynebacterium and Corynebacterium krusei are hard-to-cultivate bacteria, there are problems such as long...

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  • Multiplex quantitative PCR (Polymerase Chain Reaction) primer, probe, kit and detection method for corynebacterium bacteria and Klebsiella corynebacterium
  • Multiplex quantitative PCR (Polymerase Chain Reaction) primer, probe, kit and detection method for corynebacterium bacteria and Klebsiella corynebacterium
  • Multiplex quantitative PCR (Polymerase Chain Reaction) primer, probe, kit and detection method for corynebacterium bacteria and Klebsiella corynebacterium

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Embodiment 1

[0034] 1. Genomic DNA extraction:

[0035] 1.1 Reagents, Consumables and Instruments

[0036] 1.1.1 Reagents and consumables: Genomic DNA extraction kit (TIANamp Genomic DNA Kit, DP304-02; Tiangen Company) contains: Buffer GA; Lysis Solution GB; Buffer GD; Washing Solution PW; Elution Buffer TE; Proteinase K (20mg / mL); adsorbed; collection tube (2mL); 1.5mL sterile collection tube, absolute ethanol.

[0037] 1.1.2 Instruments: Eppendorf 5430R centrifuge, constant temperature water bath (Huanyu Technology Instrument, Jintan City, Jiangsu Province), oscillator (Dalong Xingchuang Company), Nanodrop 2000 micro-ultraviolet spectrophotometer (Thermo Company, USA).

[0038] 1.2 Preparation of human breast pus total DNA template to be tested

[0039] 1.2.1 Take the pus in the patient's breast with a disposable swab or a disposable syringe, and use Tiangen's blood genomic DNA extraction kit (the main components are: buffer GA; lysate GB; buffer GD; rinse solution PW ; Elution buffer T...

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Abstract

The invention discloses a multiplex quantitative PCR (Polymerase Chain Reaction) primer, a probe, a kit and a detection method for corynebacterium bacteria and Klebsiella corynebacterium. Aiming at corynebacterium bacterium 23S rRNA genes and Klebsiella corynebacterium 16S rRNA gene sequences, specific primers and probes are designed, whether the corynebacterium bacteria and Klebsiella corynebacterium exist in a biological sample can be identified by utilizing multiplex quantitative PCR. The primers and probes have an important guiding role in clinical targeted selective use of antibiotics. Due to reasonable usage of the antibiotics, the course of granulomatous mastitis can be effectively shortened, the condition that breast tissues are removed during the operation is avoided, physical and psychological trauma of a patient can be greatly alleviated, and an etiological basis is provided for clinical diagnosis and treatment of the granulomatous mastitis.

Description

Technical field: [0001] The invention belongs to the fields of molecular biology and medicine, and in particular relates to a multiple fluorescent quantitative PCR primer, probe, kit and detection method for bacteria belonging to the genus Corynebacterium and Corynebacterium krusei. Background technique: [0002] In recent years, in clinical practice in hospitals, it has been found that the incidence of non-lactating mastitis has gradually increased, even exceeding that of lactating mastitis. Granulomatous mastitis is an important non-lactating mammary gland inflammatory disease. Its clinical manifestations are mammary gland mass, abscess sinus tract or ulcer. The etiology of granulomatous mastitis is complex, involving local trauma, autoimmunity, bacterial infection and other factors. In recent years, more and more domestic and foreign studies have shown that Corynebacterium infection plays an important role in the development of granulomatous mastitis lesions. Recently, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/15
CPCC12Q1/6851C12Q1/689C12Q2600/16C12Q2563/107C12Q2537/143C12Q2531/113
Inventor 张亮马健周伟平李泌周香城王颀于海静王波
Owner GUANGDONG WOMEN & CHILDREN HOSPITAL
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