Fluorescence quantitative PCR (polymerase chain reaction) universal premier for detecting pathogenic aspergillus, detection probe and kit

Abstract

The invention discloses a fluorescence quantitative PCR (polymerase chain reaction) universal premier for detecting aspergillus, detection probes and a detection kit, wherein the detection kit comprises universal premiers displayed by base compositions such as SEQ ID. NO: 1 and SEQ ID. NO: 2; and the detection probe is CY5-TAAAGTTGGGTGTCGGCTGG-BHQ aiming at aspergillus fumigatus, the detection probe is FAM-TTGATTTGCGTTCGGCAAGC-BHQ aiming at aspergillus flavus, the detection probe is HEX-ACAAGTTGCAAATAAATGCGTCG-BHQ aiming at aspergillus, and / or the detection probe is ROX-ATGGTTGGAAAACGTCGGCA-BHQ aiming at aspergillus Niger. The multiple PCR detection method, premier, the probes and the kit thereof provided by the invention aim at four main pathogenic aspergilli and have high specificity and sensitivity; and the detection method is rapid, simple and convenient, and can be used for detecting and identifying the strain of pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fluorescent quantitative PCR general primer, a detection probe and a kit for detecting pathogenic aspergillus. Background technique [0002] In recent years, with the wide application of broad-spectrum antibiotics and immunosuppressants and the increasing number of immunocompromised populations such as hematological malignancies and organ transplants, the incidence of deep fungal infections is rising sharply. Among them, Aspergillus has become an important pathogenic fungus second only to Candida albicans in immunocompromised patients. Aspergillus infection occurs in up to 50% of immunocompromised patients. Aspergillus infection, especially invasive aspergillosis, has a poor prognosis. Although new antifungal drugs are emerging and used in clinical treatment, the fatality rate is still as high as 50% to 90%. Due to the non-specific clinical manifestations of the disea...

AI Technical Summary

Problems solved by technology

However, these methods have certain defects and limitations: traditional culture and microscopic examination methods have a low positive rate and take a long time; histopathological examination is an invasive operation, it is not easy to obtain materials, and immunohistochemical methods are required to further identify the bacteria species ; The Halo sign seen on imaging examinations is a characteristic manifestation of early invasive pulmonary aspergillosis, but lacks specificity; serological examination of specific antigens or metabolites such as detection of galactomannan antigen (GM antigen) and (1→3)2β2D2 glucan (G test) is quick and easy, but its sensitivity, specificity and value in early diagnosis and disease monitoring are still unsatisfactory, and the strains of Aspergillus cannot be identified

The advantage of this method is that it is simple and easy to implement, and the main disadvantage is that the specificity is not strong. The dye can bind to any double-stranded DNA, and it cannot distinguish primer dimers, non-specific amplification products and specific amplification products.

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