二代测序技术在外源性眼内炎患者病原微生物检测中的应用  

Application of metagenomics second-generation sequencing technology in detecting pathogenic microorganisms in patients with exogenous endophthalmitis

作　　者：郝昕蕾 张依 李雪杰 金玮[1] 杨安怀[1] HAO Xinlei;ZHANG Yi;LI Xuejie;JIN Wei;YANG Anhuai(Eye Center of Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China)

机构地区：[1]武汉大学人民医院眼科中心,湖北省武汉市430060

出　　处：《眼科新进展》2021年第8期750-754,共5页Recent Advances in Ophthalmology

基　　金：湖北省自然科学基金面上项目(编号2019CFB489)。

摘　　要：目的探讨二代测序技术在外源性眼内炎患者病原微生物检测中的应用。方法对2018年11月至2019年11月于武汉大学人民医院眼科中心就诊的21例(21眼)外源性眼内炎患者的临床特征和病原学结果进行分析。眼外伤史14例,内眼手术史7例。所有患者入院时均已完善相关检查并接受手术,术中尽可能清除前房及玻璃体内脓性病灶,并留存病灶样本(前房标本2份,玻璃体标本19份),通过宏基因组二代测序技术检测病原体和病原体培养两种方法对标本进行检测。对患者入院视力和末次随访视力行配对样本的非参数检验。对二代测序和病原体培养两种方法检出病原体阳性率差异行卡方检验。结果14例外源性眼内炎患者可观察到前房积脓或玻璃体内脓性病灶;21例患者中,20例患者眼内感染均得到有效控制,眼球得以保留。与入院视力相比,患者末次随访视力提高,差异有统计学意义(P=0.006)。二代测序技术检测病原体阳性率为95.24%(20/21),病原体培养阳性率为42.86%(9/21),二者差异有统计学意义(P<0.001)。病原体培养阳性的9例标本均检出单种病原体;利用二代测序方法检测,13例标本检出单种病原体,7例标本检出两种病原体。结论结合使用二代测序技术与传统的培养手段,利于检出外源性眼内炎中病原微生物,指导临床诊疗。Objective To explore the clinical application of metagenomics second-generation sequencing in detecting pathogenic microorganisms in patients with exogenous endophthalmitis.Methods Twenty-one patients(21 eyes)diagnosed with exogenous endophthalmitis at the Eye Center of Renmin Hospital of Wuhan University from November 2018 to November 2019 were selected in this study.Their clinical characteristics and etiological results were analyzed.Among them,14 patients had a history of ocular trauma,and 7 patients had a history of intraocular surgery.All patients completed necessary examinations at the time of admission and underwent surgery.During surgery,the purulent lesions in the anterior chamber and vitreous chamber were removed as much as possible,and 21 lesion specimens(2 from the anterior chamber and 19 from the vitreous chamber)were retained for detection by metagenomics second-generation sequencing and pathogen culture.A paired non-parametric test was performed for the visual acuity of patients at admission and at the last follow-up.A chi-square test was performed for the difference in the positive rate of pathogens detected by metagenomics second-generation sequencing and pathogen culture.Results Hypopyon or purulent lesions in the vitreous chamber were observed in 14 patients with exogenous endophthalmitis.Twenty patients’intraocular infection was effectively controlled,with eyeballs preserved.Compared with that at admission,the visual acuity at the last follow-up improved with statistical significance(P=0.006).The positive rate of pathogen detected by the NGS was 95.24%(20/21),and that detected by the pathogen culture was 42.86%(9/21).There was a significant difference between the two methods(P<0.001).A single pathogen was detected in all 9 positive specimens from the pathogen culture.While for specimens obtained by the NGS,a single pathogen was detected in 13 specimens and two pathogens were detected in 7 specimens.Conclusion The combined use of the NGS and conventional culture could help detect pathogenic m

关 键 词：外源性眼内炎 宏基因组测序/二代测序 病原体培养 玻璃体液/房水 

分 类 号：R771.2[医药卫生—眼科]