Development And Preliminary Clinical Application Of Ultrafiltration Membrane Pulmonary Aspergillus Assay

OBJECTIVE:Pulmonary aspergillosis as a serious invasive fungal infectious disease,there is a lack of clinical detection of Aspergillus with low detection cost,simple operation,high sensitivity,high specificity and rapid and accurate detection method.In this study,we propose to establish an ultrafiltration membrane filtration technique to detect Aspergillus,and investigate the value of ultrafiltration membrane filtration assay in patients with pulmonary Aspergillus infection by comparing the diagnostic efficacy of traditional methods(sputum culture,alveolar lavage fluid culture,bronchial brush check),serum and BALF Aspergillus antigen assay(GM test),and m NGS assay in the same batch of clinical specimens.METHODS:1.Firstly,an ultrafiltration membrane(a fiber filter membrane with a pore size of 2μm)was designed and manufactured to intercept Aspergillus and other impurity cells such as bacteria according to their size differences,and Aspergillus aggregated on the nanomicroporous filter membrane diaphragm,and then pathological staining was performed directly on the diaphragm,followed by microscopic observation.2.According to the inclusion and exclusion criteria,36 patients with Aspergillus pulmonarius infection and 49 patients with other lung diseases other than Aspergillus pulmonarius infection were finally included,and the patients’ basic information,final diagnosis,laboratory test results and results of the ultrafiltration membrane Aspergillus pulmonarius assay were collected.The sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,accuracy rate and Kappa value of the various tests were calculated,and the statistical differences between the ultrafiltration membrane Aspergillus pulmonary test and other tests were compared to evaluate the best diagnostic strategy.RESULTS:The sensitivity of the ultrafiltration membrane Aspergillus pulmonarius assay for the diagnosis of pulmonary aspergillosis was 61.1% and the specificity was 100%.m NGS assay for the diagnosis of pulmonary aspergillosis had a sensitivity of 83.3% and a specificity of 95.9%.The sensitivity of sputum culture for the diagnosis of pulmonary aspergillosis was 5.5%,and the specificity was 100%.The sensitivity of alveolar lavage fluid culture for the diagnosis of pulmonary aspergillosis was 8.3%,and the specificity was 100%.The sensitivity of serum GM assay for the diagnosis of pulmonary aspergillosis was 13.8%,and the specificity was 97.9%.The sensitivity of alveolar lavage GM test for the diagnosis of pulmonary aspergillosis is 33.3%,and the specificity is93.8%.The sensitivity of the bronchial brush test method for the diagnosis of pulmonary aspergillosis was 5.2%,and the specificity was 100%.The sensitivity of ultrafiltration membrane Aspergillus pulmonarius assay for the detection of pulmonary aspergillosis was higher than that of sputum culture(61.1% vs.8.3%,p < 0.05),serum GM(61.1% vs.13.8%,p < 0.05),alveolar lavage fluid GM(61.1% vs.33.3%,p = 0.001),and bronchial brush test(61.1% vs.5.2%,p < 0.05),and lower than that of m NGS(61.1% vs.83.3%,p=0.011).Sensitivity of different assays for detection of Aspergillus pulmonarius:m NGS > ultrafiltration membrane Aspergillus pulmonarius assay > alveolar lavage fluid GM > serum GM > BALF culture = bronchial brush test > sputum culture.Comparison of the possibility of excluding Aspergillus pulmonarius infection in this study: m NGS >ultrafiltration membrane Aspergillus test > alveolar lavage fluid GM > serum GM >bronchial brush test = BALF culture > sputum culture.The ultrafiltration membrane pulmonary aspergillus assay was highly consistent compared to clinical diagnosis(Kappa value = 0.644).CONCLUSION:1.A detection technique with high sensitivity,high specificity and reliable results was established for the detection of Aspergillus in alveolar lavage fluid.2.The ultrafiltration membrane Aspergillus pulmonarius assay has good application in patients with pulmonary Aspergillus infection.The sensitivity and accuracy of ultrafiltration membrane Aspergillus test is higher than sputum culture,serum GM,alveolar lavage fluid GM,and bronchial brush test,and it is highly consistent with the results derived from the final diagnosis in the clinic3.The ultrafiltration membrane assay has the advantages of simplicity,efficiency and accuracy,which can meet the needs of clinical diagnosis of Aspergillus infection.The detection process does not require special instruments and can be carried out in ordinary clinical laboratories,which is expected to become a new routine detection method for Aspergillus infections.