宏基因二代测序技术对脊柱感染的诊断效率及预后的影响

Diagnostic efficiency of metagenomic next-generation sequencing on spinal infection and prognosis

目的:脊柱感染是一种比较少见且治疗棘手的感染性疾病。由于经验性使用抗生素使多重耐药菌感染发生率增加,同时检测技术进步导致检出率增加,因此脊柱感染的发病率呈上升趋势。传统检测方法诊断脊柱感染存在敏感性低和耗时长等局限性。在脊柱感染的临床诊疗中,判断病原体的类型从而有针对性地使用抗生素一直是重点和难点。许多患者在脊柱感染早期因传统检测方法的局限性不能得到快速精准的诊断,造成诊断延迟,错过最佳治疗时间,给患者带来灾难性后果。临床上急需一种高特异度、高灵敏度、耗时少的检测脊柱感染病原体的技术。宏基因二代测序技术(metagenomic next-generation sequencing,m NGS)是近几年新兴的前沿技术,其可以检测样本中已知的所有病原体,已应用于诊断临床上非典型的、罕见的感染性疾病。本研究旨在分析mNGS技术在脊柱感染术后诊断病原体中的敏感度,以及其对预后的影响。方法:回顾性分析2019年1月至2021年12月中南大学湘雅医院收治的82例脊柱感染患者的临床资料,术前检测其外周血C反应蛋白(C-reactive protein,CRP)、红细胞沉降率(erythrocyte sedimentation rate,ESR)、降钙素原(procalcitonin,PCT),在术中取得患者病灶组织标本,并进行微生物培养、组织病理学检查和mNGS检测。根据mNGS检测结果,将82例患者分为针对用药组(n=71)和经验用药组(n=11)。术后定期随访,根据病理学及病原学检查结果,评价m NGS检测脊柱感染病原体的敏感度及其对预后的影响。结果:mNGS检测脊柱感染病原体的阳性率(86.59%,71/82)显著高于微生物培养(18.99%,15/79)和PCT检测(30.23%,13/43)。针对用药组与经验用药组的术前体温、白细胞计数、中性粒细胞比值和入院视觉模拟评分差异均无统计学意义(均P>0.05);术前是否使用抗生素对mNGS检测结果和微生物培养结果阳性率无明显影响(P=0.681)。针对用药组患者的术后CRP、ESR呈下降趋势;术后30 d,针对用药组的ESR较经验用药组明显下降(P=0.044)。结论:相比于微生物培养和PCT检测,mNGS检测脊柱感染病原体的敏感度更高;与经验用药相比,根据mNGS检测结果针对性使用抗生素的患者预后更好。

Objective:Spinal infection is a rare infectious disease that is difficult to treat.The incidence of spinal infection is on the rise with the experiential use of antibiotics,the increasing incidence of drug-resistant bacteria,and the improvement of detection techniques.Traditional detection methods have limitations such as low sensitivity and long time-consuming in the diagnosis of spinal infection.In the clinical diagnosis and treatment of spinal infection,it has always been the focus and difficulty to determine the type of pathogens and to use antibiotics in a targeted manner.Many patients in the early stage of spinal infection due to the limitations of traditional detection methods cannot be quickly and accurately diagnosed,resulting in diagnosis delay,missed the best treatment time,bringing disastrous consequences to patients.There is an urgent need for a high-specificity,high-sensitivity,and time-saving test technique in clinical practice,which can simultaneously distinguish and identify the pathogen of spinal infection.Metagenomic next-generation sequencing(mNGS)is a new frontier technology emerging in recent years.It can detect all known pathogens in samples and has been used to diagnose clinically atypical and rare infectious diseases.This study aims to analyze the sensitivity of m NGS technique in diagnosing pathogens after spinal infection and its effect on prognosis.Methods:Clinical data of 82 patients with spinal infection admitted to Xiangya Hospital of Central South University from January 2019 to December 2021 were retrospectively analyzed.Peripheral blood erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),and procalcitonin(PCT)were routinely performed before surgery,and focal tissue specimens were obtained during surgery.Microbial culture,histopathological examination,and m NGS detection were performed.All patients were assigned into a targeted medication group(n=71)and an experienced treatment group(n=1)based on the results of mNGS.After regular follow-up,the sensitivity of mNGS to detect pathogens of spinal infection and its effect on prognosis were evaluated.Results:The positive rate of mNGS(86.59%,71/82)was significantly higher than that of microbial culture(18.99%,15/79)and PCT(30.23%,13/43).There were no significant differences in preoperative temperature,white blood cell count,neutrophil ratio,and scores of Visual Analogue Scale between the targeted medication group and the experienced treatment group.Preoperative use of antibiotics had no significant effect on the positive rate of mNGS and microbial culture(P=0.681).According to the targeted medication group,postoperative CRP and ESR showed a decreasing trend,and the ESR was significantly lower than that of the experienced treatment group at 30 days follow-up(P=0.044).Conclusion:Compared with the microbial culture or PCT,mNGS has a higher sensitivity rate to detect pathogens of spinal infection.Patients receiving targeted antibiotics based on the results of mNGS have better outcomes than those receiving the experienced medicine.