二代测序技术在免疫缺陷并发重症肺炎患者病原体检测中的应用

Application of NGS technology in pathogen detection of patients with immunodeficiency complicated with severe pneumonia

目的探讨二代测序技术(NGS)在免疫缺陷并发重症肺炎患者病原体检测中的应用价值,为临床工作提供参考。方法前瞻性选取2018年11月—2022年5月晋江市医院100例免疫缺陷并发重症肺炎患者,采集肺泡灌洗液,均进行传统病原学及NGS检测,比较两种方法检测差异,并分析影响NGS检测结果的相关因素。结果100例免疫缺陷并发重症肺炎患者NGS阳性检出率(84.00%)高于传统病原学阳性检出率(40.00%)(P<0.05);NGS检出病原体179株,传统病原学检查检出病原体71株。NGS对真菌及病毒的检出率明显高于传统病原学检测,差异有统计学意义(P<0.05);在病原体种类数方面,NGS检测对2种及≥3种病原体的检出率高于传统病原学检测,差异有统计学意义(P<0.05);NGS检测结果在白细胞计数(WBC)、反复感染、C反应蛋白(CRP)、中性粒细胞比例(N)、降钙素原(PCT)方面比较,差异有统计学意义(P<0.05);LASSO回归筛选出5个重要预测因素:WBC、反复感染、CRP、N、PCT;Logistic回归分析显示,WBC、反复感染、CKP、N、PCT均是免疫缺陷并发重症肺炎患者NGS检测结果的独立影响因素(P<0.05)。结论NGS对免疫缺陷并发重症肺炎患者的病原体检测效能优于传统病原学,对临床治疗有积极指导价值;WBC、反复感染、CRP、N、PCT等因素可能影响NGS检测结果。

Objective To explore the application of second generation sequencing technology(NGS)in the detection of pathogens in patients with immunodeficiency complicated with severe pneumonia,and to provide reference for clinical work.Methods 100 cases of immunodeficiency complicated with severe pneumonia in our hospital from November 2018 to May 2022 were selected.Alveolar lavage fluid was collected,and traditional etiological and NGS detection were performed.The differences between the two methods were compared,and the related factors affecting the results of NGS detection were analyzed.Results The positive detection rate of NGS in 100 patients with immune deficiency complicated with severe pneumonia was 84.00%,which was higher than that of traditional etiology 40.00%(P<0.05).179 strains of pathogens were detected in ngs,and 71 strains were detected in traditional etiological examination.The detection rate of NGS for fungi and viruses was significantly higher than that of traditional etiology detection,and the difference was statistically significant(P<0.05);In terms of the number of pathogen species,the detection rate of NGS detection for 2 and≥3 pathogens was higher than that of traditional etiology detection,and the difference was statistically significant(P<0.05).There were statistically significant differences in white blood cell count(WBC),repeated infection,C-reactive protein(CRP),neutrophil ratio(N)and procalcitonin(PCT)in NGS detection results(P<0.05).LASSO regression screened out five important predictors:WBC,repeated infection,CRP,N,and PCT.Logistic regression analysis showed that all the five predictors were independent influencing factors of NGS test results in patients with immune deficiency complicated with severe pneumonia(P<0.05).Conclusion NGS is superior to traditional etiology in detecting pathogens in patients with immunodeficiency complicated with severe pneumonia,and has positive guiding value for clinical treatment;WBC,recurrent infection,CRP,N,PCT and other factors may affect the detection results of ngs.