| The incidence rate of invasive fungal infection in hematologic malignancies has gradually increased in the past 20 years,and the invasive aspergillosis has become the second major cause in deep fungal infection behind candidasis.Early rapid and accurate diagnosis can effectively reduce the incidence of fungal resistance and it can significantly increase the survival rate of patients.Because of long-time consuming and low sensitivity of traditional fungal culture,and poor specificity of imageology assay, finding a fast and reliable detection of aspergillosis infection diagnosis has become a hot issue in clinical trials.Objective: The aim of this study was to develop and evaluate a real-time PCR diagnostic techniques for Invasive Aspergillosis (IA) in hematologic malignancies patients with designing primers . We expect to provide a rapid and accurate real-time PCR assay for the clinical early diagnosis of IA.The test will be evaluated by DNA isolation from patients who were at high risk of IA,and detect them in a single-blind manner.Methods:⑴According to the GeneBank database, we used the Blast and DNAMan tools to search conservative sequence of aspergillus niger and Aspergillus nidulans.Probes were designed.The standards were established and used to establish standard curves which were evaluated later.⑵We extracted the fungal DNA in the blood samples collected, then detected them with real-time PCR assay in a single-blind manner so as to statistically evaluate the value of their diagnosis.Results:⑴The real-time PCR standard curve was completed Y|^ = -3.538Χ+42.104,R2=0.9996(A.niger); Y|^ = - 3.380Χ+43.474,R2=0.9990 (A.nidulans),and had high specificity and reproducibility.⑵.Compared to GM assay,the real-time PCR could find the false negative,indicated that it had better sensitivity.Conclusion:The real-time PCR diagnostic techniques for invasive aspergillosis in hematologic malignancies patients was successfully established,which had high sensitivity,specificity and reproducibility.
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