Method for identifying candida glabrata by high-resolution-ratio melting curve

Abstract

The invention provides a method for identifying candida glabrata by a high-resolution-ratio melting curve, wherein the method includes the following steps: step 1, preparing DNA templates of strains to be identified and a control strain, wherein the strains to be identified includes candida glabrata, candida nivariensis and candida bracarensis; step 2, designing upstream and downstream primers; step 3, obtaining a high-resolution-ratio melting curve chart of each strain by a high-resolution-ratio melting curve technology HRM, and specifically using a Real-time PCR technology, to obtain a high-resolution-ratio dissolution curve chart of each strain through real-time detection of the change of the fluorescence signal value in the DNA melting process; step 4, analyzing and identifying the specific types of the strains to be identified by analysis software; step 5, testing the nucleoside acid sequence of the strain having the result being positive in the step 3; and step 6, verifying the sensitivity of the identification method. The method for identifying candida glabrata by the high-resolution-ratio melting curve can clearly identify different species in the candida glabrata complex,and is simple to operate, accurate and efficient.

Description

technical field [0001] The invention relates to the field of strain identification, in particular to a method for identifying Candida glabrata with a high-resolution melting curve. Background technique [0002] The Candida glabrata complex is a clinically important pathogenic fungus that causes lethal diseases including invasive fungal infections. The Candida glabrata complex includes three species of human pathogenic Candida: Candida glabrata, Candida nivariensis, and Candida bracarensis. Candida glabrata is a common pathogenic fungus clinically, its isolation rate in North America and Europe is only lower than that of Candida albicans, ranking second, and it is the fourth most common Candida species in my country; the latter two are rare Candida species Bacteria, accounting for <0.5% in yeast. According to reports, the sensitivity of the latter two rare bacteria to azoles is lower than that of Candida glabrata, so accurate identification of the bacteria is very importa...

AI Technical Summary

Problems solved by technology

[0003] The biochemical characteristics of the three bacteria in the Candida glabrata complex are similar, and the traditional clinical phenotypic identification methods (including Vitek 2Compact and API ID32C) cannot identify the two rare bacteria to the species level or misidentify them as Candida glabrata

MALDI-TOF MS is an emerging technology that plays an important role in the identification of clinical microbial species. It can correctly identify common clinical yeast species, but its identification ability depends on the database: when there is a lack of rare and emerging species in the database , often unidentified or misidentified

Vitek MS v2.0 and Bruker Biotyper v3.1, widely used mass spectrometry systems in clinical practice, cannot correctly identify two rare bacteria to the species level

Using polymerase chain reaction (PCR) to amplify the intertranscriptional region of ribosomal DNA and determine the nucleic acid sequence is the "gold standard" for species identification, but the operation is complicated and time-consuming

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