宏基因组二代测序在粒细胞缺乏伴发热患者病原菌检测中的价值

Clinical application of metagenomic next-generation sequencing for determining pathogens in febrile neutropenic patients

目的 探讨宏基因组二代测序(mNGS)技术在化疗后粒细胞缺乏伴发热患者病原菌检测中的价值。方法 回顾性分析2019年6月—2020年9月西安市中心医院19例化疗后粒细胞缺乏伴发热患者的临床资料。比较mNGS与传统微生物培养、免疫学相关检测[G试验、GM试验、革兰阴性菌脂多糖(GNBL)]的阳性率和所需时间。采用受试者工作特征(ROC)曲线分析3种方法检测病原的效能。结果 19例患者临床送检样本包括血液样本(8例)、咽拭子样本(7例)、脑脊液样本(1例)、胸腔积液和腹腔积液样本(2例)、尿液样本(1例)。19例样本mNGS、微生物培养、免疫学相关检测阳性率分别为68.4%、26.3%、63.2%。13例mNGS阳性样本中,10例检出革兰阴性菌、4例检出革兰阳性菌、4例检出真菌、3例检出病毒、1例检出支原体;mNGS、微生物培养、免疫学相关检测检出病原所需时间分别为(47.0±13.2)、(84.0±31.6)、(24.8±16.4)h,mNGS较微生物培养所需时间更短(P<0.05)。ROC曲线分析结果显示,mNGS、微生物培养、免疫学相关检测检出病原的曲线下面积分别为0.806、0.688、0.550,敏感性和特异性分别为72.10%和68.46%、30.70%和82.94%、70.9%和30.70%;3种方法比较,差异有统计学意义(P<0.05)。结论 mNGS可以提高病原检测效率和可靠性,较传统微生物培养和免疫学相关检测,在粒细胞缺乏伴发热患者病原检测中有一定的优势。

Objective To access the clinical role of metagenomic next-generation sequencing(mNGS) for determining pathogens in patients with febrile neutropenia(FN) after chemotherapy. Methods The data of 19patients who were clinically diagnosed with FN after chemotherapy and were treated in Xi’an Central hospital from July 2019 to September 2020 were collected. The positive rates and spending times of mNGS,microculture and immunologic test were compared. The efficiency of mNGS,microculture and immunologic test [G test,GM test and Gram-negative bacterium lipopolysaccharide(GNBL)] was analyzed by receiver operating characteristic(ROC)curve. Results Totally,19 patients were enrolled,and 8 blood samples,7 throat swab samples,1 cerebrospinal fluid sample,2 hydrothorax and ascites samples and 1 urine sample were collected. The positive rates of mNGS,microculture and immunologic test were 68.4%,26.3% and 63.2%,respectively. Among the 13 positive samples in mNGS,there were 10 cases of Gram negative bacteria,4 cases of Gram positive bacteria,4 cases of fungi,3 cases of viruses and 1 case of mycoplasma. The spending times of mNGS,microculture and immunologic test were(47.0±13.2) h,(84.0±31.6) h and(24.8±16.4) h,and mNGS spent shorter time than microculture(P<0.05). The sensitivities of mNGS,microculture and immunologic test were 72.10%,30.70% and 70.9%,the specificities were 68.46%,82.94% and 30.70%,and the areas under curves were 0.806,0.688 and 0.550,respectively. There was statistical significance among the 3 methods(P<0.05). Conclusions The mNGS has good efficiency,and it is reliable than microculture and immunologic test,which has advantages in determining pathogens among FN patients after chemotherapy.