Rapid diagnosis reagent kit for detecting four/more than four invasive candidiasis

A technology for rapid diagnosis and application of methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of inappropriate tissue biopsy methods, low positive rate, unacceptable patients, etc., to reduce deaths rate and sequelae, high detection sensitivity, and highly reproducible results

Inactive Publication Date: 2009-08-19
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fungal culture is affected by factors such as sampling time, sampling location, sample factors, and culture conditions. The positive rate has always been low, and it is time-consuming and laborious. Routine culture takes 3 days and cannot play a role in guiding clinical early targeted treatment.
Although the serological method is widely welcomed by laboratories due to its simple operation process, the application range of this method is narrow, and there are cross-antigens, which are prone to false positives or false negatives, and the species cannot be identified, which cannot meet the needs of diagnosis
Histopathological identification method is a method for the diagnosis of clinical invasive candidiasis, but because it is an invasive examination, it is difficult for patients to accept, and patients with severe invasive fungal infection are often accompanied by thrombocytopenia and poor coagulation function, so it is generally not suitable tissue biopsy method
The method has a low positive rate
Therefore, the aforementioned laboratory diagnostic methods limit their application in clinical diagnosis.

Method used

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  • Rapid diagnosis reagent kit for detecting four/more than four invasive candidiasis
  • Rapid diagnosis reagent kit for detecting four/more than four invasive candidiasis
  • Rapid diagnosis reagent kit for detecting four/more than four invasive candidiasis

Examples

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specific Embodiment 1

[0031] This embodiment 1 constitutes a fluorescent quantitative PCR rapid diagnostic kit for detecting four kinds of invasive candida and its application method. It includes a box body and a fluorescent quantitative PCR reaction solution arranged in the box body, positive quality control products and negative quality control products of four kinds of Candida; the fluorescent quantitative PCR reaction solution contains specific primers and fluorescent lights corresponding to four kinds of Candida Probe; The positive quality control product of described invasive Candida is the DNA sample obtained by "boiling method" from the standard bacterial strain of Candida albicans, Candida krusei, Candida glabrata, and Candida tropicalis, and the negative quality control product for sterilized triple distilled water.

[0032] The preparation method of described three kinds of substances is as follows:

[0033] 1) Preparation of fluorescent quantitative PCR reaction solution: 12.5 μl of Br...

specific Embodiment 2

[0050] Example 2 constitutes a fluorescent quantitative PCR rapid diagnostic kit for detecting four / more than four types of invasive Candida and its application method.

[0051] The characteristics of this specific embodiment 2 are: the positive quality control product of described invasive Candida is made of Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis plus several other standard bacterial strains of invasive Candida The DNA sample obtained by "boiling method", the negative quality control substance of the invasive Candida is sterilized triple distilled water. The other invasive Candida can be one or a combination of two or more of Candida parapsilosis, Candida quaternidae, Candida portuguese and Candida lactis. All the other are identical with specific embodiment 1.

[0052] In conjunction with Table 2 and Table 3, use the kit of the present invention to amplify 13 kinds of 94 clinically common pathogenic fungi, 5 kinds of bacteria and human whole ...

specific Embodiment 3

[0054] Dilute the suspension of Candida albicans, Candida krusei, Candida glabrata, and Candida tropicalis by 10 times: 5×10 8 , 5×10 7 , 5×10 6 , 5×10 5 , 5×10 4 , 5×10 3 , 5×10 2 , 5×10 1 CFU / ml. Fluorescent quantitative PCR amplification was carried out after the DNA was extracted according to the above method, and the result was 5×10 3 The "S" curve can still be observed in CFU / ml, indicating that the fluorescence quantitative PCR method can detect at least 5×10 3 CFU / ml of the above four kinds of Candida, the kit of the present invention has strong sensitivity.

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Abstract

The invention relates to a quick diagnostic kit used for testing four or more than four kinds of invasive candidas and an application method thereof. The quick diagnostic kit is characterized in that the kit is internally provided with fluorescent quantitative PCR reaction liquid containing specific primers and fluorescent probes corresponding to the candidas which are at least four kinds as well as positive control materials and negative control materials of the candidas which are at least four kinds; Ct value tested by a fluorescent quantitative PCR tester is used for judging. The kit has the characteristics of good specificity and high sensitivity; due to double control of high specificity amplification of the primers and high specificity hybridization of the fluorescent probes, the false positive is low; therefore, the invention can be used for simultaneously testing four kinds of invasive candidas which have different sensitivities for clinical first-line antifungal drugs such as fluconazole and the like in a simple, convenient, rapid, special and sensitive way.

Description

technical field [0001] The invention relates to a rapid diagnostic kit and an application method thereof, in particular to a fluorescent quantitative PCR rapid diagnostic kit for detecting four / more than four types of invasive candida and an application method thereof. It belongs to the technical field of fungal nucleic acid detection and is suitable for the rapid detection of Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis and other invasive candida in clinical and scientific research. Background technique [0002] In recent years, with the extensive development of organ transplantation and bone marrow transplantation, the increase of AIDS and cancer patients, and the extensive application of corticosteroids, broad-spectrum antibiotics, and immunosuppressants, the incidence of invasive candida infection has increased significantly. A prospective study in the United States showed that fungal infections accounted for 9.5% of nosocomial bloodstream infec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64C12R1/645
Inventor 黎毅敏杨淳黄红川刘晓青何为群袁锦屏杨灵邱桂霞张彦峰
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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