Rapid Fungus detection kit and detection method using same

A detection kit and detection method technology, applied in the field of clinical microorganism identification, can solve the problems of prolonged hospitalization, drug resistance test costs, treatment failure, etc., and achieve the effects of shortened identification time, low cost, and convenient operation.

Active Publication Date: 2011-02-16
呼和浩特迪安医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Inappropriate use of antibiotics often leads to treatment failure, resulting in prolonged hospitalization, complications, drug resistance, and increased testing costs

Method used

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  • Rapid Fungus detection kit and detection method using same
  • Rapid Fungus detection kit and detection method using same
  • Rapid Fungus detection kit and detection method using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: the preparation method of kit.

[0064] (1) DNA extraction reagents:

[0065] Buffer I is 9g / L NaCl aqueous solution;

[0066] Buffer II is 10g / L SDS, 2% (v / v) Triton X-100, 100mmol / L NaCl, 10mmol Tris-HCl (pH 8.0), 1mmol / L EDTA, and the solvent is water;

[0067] Buffer III is a saturated NaCl aqueous solution (about 6mol / L);

[0068] Buffer TE is 10mmol / L Tris-HCl (pH 8.0), 1mmol / L EDTA, and the solvent is water;

[0069] Among them: NaCl (purchased from Shanghai Shisihewei Chemical Co., Ltd.), SDS (purchased from Sigma Company in the United States), EDTA (purchased from Hangzhou Chemical Reagent Co., Ltd.), Tris-HCl (purchased from Sigma Company in the United States), Tris-HCl (Tris-base was purchased from Sigma Company in the United States), hydrochloric acid purchased from Hangzhou Chemical Reagent Co., Ltd.), Triton X-100 (purchased from Sigma, USA)

[0070] (2) Reaction solution:

[0071] PCRBuffer: 0.1% (v / v) NP-40 (purchased from Sigma, USA),...

Embodiment 2

[0088] Embodiment 2: detection method.

[0089] Instruments: Bio-Rad S1000 PCR instrument, Beckman Microfuge 22R desktop micro-centrifuge, Beijing Liuyi agarose gel electrophoresis instrument, Shanghai Peiqing gel imaging system, QIAGEN PyroMark Q96ID sequencer.

[0090] (1) Extracting fungal DNA, taking clinical urine samples as an example, specifically includes the following steps:

[0091] (1a) Take 2ml of clinical urine sample in a centrifuge tube and centrifuge at 12000rpm for 10min at high speed;

[0092] (1b) Discard the supernatant, add 800ul Buffer I, and vortex for 30 seconds;

[0093] (1c) 12000rpm high-speed centrifugation for 5min;

[0094] (1d) Discard the supernatant, invert and blot dry as much as possible, add 300ul Buffer II, and shake on a high-speed oscillator for 10min;

[0095] (1e) Add 300ul Buffer III to the centrifuge tube and vortex for 30s;

[0096] (1f) 4000rpm centrifugal 15min;

[0097] (1g) Put the supernatant obtained in (1f) into another c...

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Abstract

The invention relates to the field of clinical microorganism identification, and provides a rapid fungus detection kit and a detection method using the same. The kit comprises two universal primers for amplifying an internal transcribed spacer (ITS1) of a fungus 18S, 5.8S, a sequencing primer for determining a sequence of a reverse section of the ITS1, a magnetic bead marked by an avidin, and a sequencing result and database comparison and judgment species. By using the kit, the fungus in a clinical sample can be distinguished by one-time sequencing, so the kit can be used for clinical diagnosis. The kit not only gains precious rescue time for clinical diagnosis and treatment, but also has the advantages that: the cost is low, the kit is convenient to operate and the specificity is high.

Description

technical field [0001] The invention belongs to the technical field of clinical microorganism identification, and in particular relates to a kit and a detection method for rapidly identifying fungi in clinical samples. Background technique [0002] With the wide application of tumor radiotherapy, chemotherapy, broad-spectrum antibiotics and immune agents, systemic and opportunistic fungal infections are increasing, especially in hematologic malignancies, tumors, bone marrow transplantation and organ transplantation. For example, 18% to 50% of patients have invasive fungal infection after bone marrow transplantation; the prevalence of fungal infection in organ transplant recipients and malignant tumor patients is as high as 20% to 40%. According to the analysis of my country's nosocomial infection detection network, nosocomial fungal infections rose from 13.9% in the early 1990s to 17.1% to 24.4% in the late 1990s. With the increasing number of fungal infections, the use of a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 任绪义张玉娥虞闰六
Owner 呼和浩特迪安医学检验所有限公司
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