Prophylaxis Of Invasive Fungal Infection With Different Administration Regimens Of Itraconazole In Patients With Acute Myeloid Leukemia: A Report From A Randomized, Controlled Trail

[Background]:The morbidity of invasive fungal infections (IFI) in patients with hematological malignancy is increasing these years. And IFI is one of the leading causes of mortality and morbidity. Because of the difficulty of early diagnosis, high cost of therapy, and high mortality of IFI, prophylaxis of IFI in high risk patients are drawing more and more attention. Itraconazole is a triazole antifungal agent with wide antifungal spectrum, less adverse events and relatively cheaper than other antifungal agents, so it is fit for antifungal prophylaxis.[Purpose]:1. To evaluate the effect of antifungal prophylaxis of itraconazole in patients with acute myeloid leukemia (AML).2. To evaluate the safety of antifungal prophylaxis of itraconazole in patients with AML.3. To evaluate the correlation of the antifungal effect and the adverse events with the serum concentration.[Methods]:1. Eligibility Patients were randomized separately according to different chemotherapy of induction and consolidation.2. Patients in oral group were administrated with itraconazole oral solution 2.5mg/kg bid (7am,7pm). Patients in injection/oral group were administrated with itraconazole injection 200mg q12h for the first two days, then oral solution 2.5mg/kg bid for the next period.3. Blood samples were collected in different time for measurement of the serum itraconazole level.4. The morbidity of IFI and the adverse events were observed.5. For categorical variables, the chi-square statistic test was used, t test was used for mean comparison of two groups, p value of<0.05 was regarded as significant.[Results]:1. The morbidity of IFI in injection/oral group and oral group were 10.1% and 20.9%(p=0.0095), respectively. The morbidity of adverse events in both group were fairly low, and the difference between these two groups was of no significance.2. The serum itraconazole level of injection/oral group and oral group were (698±399) ng/ml and (573±363) ng/ml (P<0.01), respectively. The serum itraconazole levels of patients who developed IFI were significantly lower than those of patients who did not develop IFI.[Conclusion]:1. Antifungal prophylaxis with itraconazole in patients with AML was effective and safe.2. Prophylactic effect with injection/oral itraconazole was superior to that with itraconazole oral solution, While the adverse events did not increase with injection/oral itraconazole.3. Prophylactic effect was firmly correlated with the serum itraconazole level. Monitoring of the serum itraconazole level and adjusting itraconazole administration dose accordingly should be conducted during the therapy course. [Objective]To establish a novel post-transplant assay approach for quantitative assessment of mixed hematopoietic chimerism using real-time polymerase chain reaction (PCR) of single nucleotide polymorphisms (SNPs), and analyse the application of real-time SNP-PCR method for patients treated by allogeneic HSCT.[Method]For the purpose of developing an chimerism analysis method for Chinese patients which underwent the hematopoietic stem cell transplantation, we searched the dbSNP database and got seven SNPs to development an informative panel. Establish a SNP——based quantitative method using real-time PCR, and then analyse three clinical samples received the patients who had relapsed after allogeneic stem cell transplantation.[Result]1. Selection of the SNP panelWe searched and got seven SNPs from dbSNP database. These seven SNPs include three type of base substitution, in which 3 of A/G,3 of C/T, lof T/G type. The selection guideline is that the allele frequence of each SNP should be between 0.3?0.8 and all SNPs should distributed on th different chromosomes. For generalizing the detection method, we made the efforts to cover the various base substitution type.2. Establish a SNP-based quantitative method using real-time PCR.In order to detect the donor/recipient chimerism we produced standard curves of all SNP locus. The negative and positive template were validated by repeated genotyping. Then we made the gradient concentration ranged from 100 to 0.01%. It showed that the linear relationship of this standard curve was fine.0.01%. For example, rs4245, after amplifying this set of templates with real—timePCR, the operation software output standard curve:y=-3.467x+27.722. The amplification efficiency was 0.94 and R2was 0.998. The sensitivity this standard curve reached was not lower than 0.01 %. and the amplification effect of the standard sample template is also good.3. Analysis of the clinical samplesThe percentages of recipient cell are 16.23%,55.86% and 89.41% respectively. It shows the prognostic value of SNP-PCR method for relapse. [Conclution]We established the quantitative detection method by preparation of the artificial chimerism, and this method made a greater improvement in detection sensitivity, accuracy and stability than traditional methods.