The role of four-generation Oxford nanopore sequencing technology in searching for pathogenic bacteria in peripros-thetic infection
支力强 1常晓峰 2马建兵 1王朝阳 3昝强 3曹士航 3孙相祥1
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作者信息
1. 西安市红会医院关节外科,西安 710021
2. 西安市红会医院关节外科,西安 710021;西安医学院研究生工作部,西安 710021
3. 陕西中医药大学附属医院骨关节科,咸阳 712000
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摘要
目的 探讨纳米孔测序技术(Oxford nanopore technologies,ONT)在假体周围感染(periprosthetic joint infec-tion,PJI)病原菌确定中的应用价值.方法 收集2021年10月至2023年3月于西安市红会医院关节外科收治的符合美国骨骼肌肉感染协会(Musculoskeletal Infection Society,MSIS)2018年PJI诊断标准的患者32例(PJI组),男15例、女17例,年龄(63.93±8.93)岁;收集同期不符合2018年MSIS的PJI标准的翻修患者32例作为对照(非PJI组),男13例、女19例,年龄(65.53±8.54)岁.所有患者均在术前或术中穿刺抽取关节液,标本分别进行ONT、宏基因组二代测序(metagenomic next generation sequencing,mNGS)检测及微生物培养检测.对两组绘制三种检测技术的受试者工作特征曲线(receiver operat-ing characteristic,ROC),计算并比较灵敏度、特异度、阳性预测值、阴性预测值及约登指数以评价不同检测方法在PJI中的检测效能.结果 32例PJI组中ONT阳性30例,共检出30种病原菌,检测时间为(22.37±8.36)h;mNGS阳性31例,共检出33种病原菌,检测时间为(46.25±9.36)h;微生物培养阳性17例,共检出8种病原菌,检测时间为(96.23±15.62)h.32例非PJI组中ONT、mNGS阳性分别为1和5例,分别检出1种和3种病原菌,微生物培养结果均为阴性.ONT与mNGS的检测时间及曲线下面积(area under the curve,AUC)分别为(22.37±8.36)h、0.953[95％CI(0.901,1.006)]和(46.25±9.36)h、0.906[95％Cl(0.835,0.977)],均优于微生物培养的(96.23±15.62)h、0.766[95％CI(0.678,0.853)],差异有统计学意义(P＜0.05).ONT、mNGS、微生物培养的灵敏度分别为0.938、0.969、0.531,特异度分别为0.969、0.844、1.000,约登指数分别0.906、0.813、0.531.结论 ONT检测在PJI诊断过程中较mNGS和微生物培养具有更高的诊断效能,在检出时间上也更有优势,也提示部分PJI并非单一微生物感染所致.
Abstract
Objective To explore the application value of Oxford nanopore technologies(ONT)in the diagnosis and treat-ment of periprosthetic joint infection(PJI).Methods A prospective analysis was conducted on 32 patients with PJI admitted to the joint department of Xi'an Honghui Hospital from October 2021 to March 2023,who met the 2018 PJI diagnostic criteria of the American Skeletal Infection Society(MSIS),including 15 males and 17 females with an average age of 63.93±8.93 years.32 revi-sion patients who did not meet the 2018 MSIS PJI criteria during the same period were collected as controls(non PJI group),in-cluding 13 males and 19 females with an average age of 65.53±8.54 years.All patients underwent joint fluid puncture before or during surgery,and the specimens were tested by ONT,metagenomic next generation sequencing(mNGS),and general microbial culture.The receiver operating characteristic(ROC)curves were drawn for both groups,and the sensitivity,specificity,positive predictive value,negative predictive value,and Youden index of the three detection techniques were calculated and compared to evaluate the detection efficiency of different detection methods in PJI.Results Among the 32 patients with PJI,30 were positive for ONT,with a total of 30 pathogenic bacteria detected,and the detection time was 22.37±8.36 h.31 were positive for mNGS,with a total of 33 bacterial species detected,and the detection time was 46.25±9.36 h.17 were positive for microbial culture,with a total of 8 bacterial species detected,and the detection time was 96.23±15.62 h.Among the 32 patients with non PJI group,1 was positive for ONT and 5 were positive for mNGS,with a total of 1 and 3 bacterial species detected,respectively.The results of mi-crobial culture were all negative.The detection time and area under the curve(AUC)of ONT and mNGS were 22.37±8.36 h and 0.953[95％CI(0.901,1.006)],46.25±9.36 h and 0.906[95％CI(0.835,0.977)],respectively,which were better than those of mi-crobial culture 96.23±15.62 h and 0.766[95％CI(0.678,0.853)],and the difference was statistically significant(P＜0.05).The sen-sitivity of ONT,mNGS,and microbial culture were 0.938,0.969,and 0.531,respectively,and the specificity was 0.969,0.844,and 1.000,respectively.The Jordan index was 0.906,0.813,and 0.531,respectively.Conclusion ONT testing has higher diagnostic efficacy than mNGS and microbial culture in the diagnosis of PJI,and also has advantages in detection time.It also suggests that some PJI are not caused by a single microbial infection.
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