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角膜移植术后植片失功患者的病毒学检测

Virological analysis of patients with corneal graft failure after keratoplasty
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摘要 目的调查角膜移植术后植片失功患者再次行角膜移植手术时的病毒学检测结果,以及多种病毒检测手段的诊断效能。方法采用病例对照研究,连续纳入2018年3—12月于北京大学第三医院眼科就诊的植片失功患者14例作为植片失功组,同时连续纳入大泡性角膜病变(BK)患者15例作为BK组,所有患者均为单眼发病且具有角膜移植手术指征。手术当日取静脉血获得血清样本,角膜移植术中获得房水及角膜组织标本。采用PCR法检测房水、角膜标本中病毒DNA,酶联免疫吸附试验检测血清及房水中病毒抗体含量,并计算Goldmann-Witmer系数(GWC)。检测病毒种类包括单纯疱疹病毒(HSV)、带状疱疹病毒(VZV)和巨细胞病毒(CMV)。植片失功组是否存在高眼压表现、既往多次角膜移植手术史及病毒性角膜炎病史与任一病毒检测结果阳性间的关系采用kappa一致性检验分析。结果植片失功组本次角膜移植术中病毒检测共9眼(9/14)至少一项病毒检测结果阳性,其中CMV阳性6眼(6/14),VZV阳性3眼(3/14);房水PCR阳性5眼(5/14),角膜组织PCR阳性5眼(5/14),房水病毒抗体GWC升高3眼(3/10),病毒DNA与抗体检测的一致性较差。BK组检测发现2眼(2/15)房水CMV GWC系数升高。植片失功组病毒检测阳性率为64.2%(9/14),明显高于BK组的13.3%(2/15),差异有统计学意义(P=0.014)。植片失功组中7眼(7/14)有高眼压表现、3眼(3/14)有多次角膜移植手术史、6眼(6/14)既往有病毒性角膜炎病史,但以上因素均与术中病毒检测结果一致性差(kappa=0.143、-0.155、-0.286)。结论病毒感染为植片失功的常见病因,再次行角膜移植术中联合多种病毒检测手段有助于明确病因及病毒感染状态,指导后续治疗。 Objective To investigate the virological testing results of patients with corneal graft at the time of repeat keratoplasty and the diagnostic efficacy of multiple viral examinations.Methods A case-control study was conducted.A total of 14 consecutive patients diagnosed with corneal graft failure were enrolled as graft failure group from March 2018 to December 2018 in Peking University Third Hospital,and 15 consecutive patients diagnosed with bullous keratopathy(BK)were enrolled as BK group in the meantime.All patients had unilateral involvement and indications for keratoplasty.Serum samples were collected from venous blood on the day of surgery,and specimens of aqueous humor and corneal tissue were obtained during corneal transplantation.Viral DNA in aqueous humor and corneal specimens was detected by real-time polymerase chain reaction(PCR).The level of viral antibodies in serum and aqueous humor was determined by enzyme-linked immunosorbent assay and the Goldmann-Witmer coefficient(GWC)was calculated.The tested viral species included herpes simplex virus(HSV),herpes zoster virus(VZV),and cytomegalovirus(CMV).For graft failure group,the relevance between elevated intraocular pressure,multiple previous keratoplasty surgeries,histories of viral keratitis and any positive result of viral analyses in this study were measured by the kappa consistency test.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University Third Hospital(No.2017299-02).Written informed consent was obtained from each subject.Results At the time of keratoplasty,9 out of 14 eyes in the graft failure group tested positive for at least one type of virus,with 6 eyes positive for CMV and 3 eyes positive for VZV.Positive aqueous humor PCR analysis detected VZV in 5 out of 14 eyes.Corneal tissue PCR analysis detected CMV in 5 out of 14 eyes.Positive GWC calculations detected CMV in 3 out of 10 eyes.The concordance between viral DNA and antibody detection was poor.All eyes in BK group were negative for the virological test,except for 2 eyes(2/15)with elevated aqueous humor GWC for CMV.The prevalence of viral infection was 64.2%(9/14)in graft failure group,which was significantly higher than 13.3%(2/15)in BK group(P=0.014).In graft failure group,7 eyes(7/14)had elevated intraocular pressure,3 eyes(3/14)had multiple keratoplasty surgeries,and 6 eyes(6/14)had viral keratitis before this keratoplasty.However,none of these factors showed significant relevance with positive virological results(kappa=0.143,-0.155,-0.286).Conclusions Viral infection has become a major cause of corneal graft failure.A combination of various virological analyses during keratoplasty can help to clarify the etiology and the viral infection status,and ultimately guide subsequent treatment.
作者 郭雨欣 孙彬佳 彭荣梅 洪晶 Guo Yuxin;Sun Binjia;Peng Rongmei;Hong Jing(Department of Ophthalmology,Peking University Third Hospital&Beijing Key Laboratory of Restoration of Damaged Ocular Nerve,Beijing 100191,China;Eye Hospital and School of Ophthalmology and Optometry,Wenzhou Medical University,Wenzhou 325027,China)
出处 《中华实验眼科杂志》 CAS CSCD 北大核心 2023年第10期985-990,共6页 Chinese Journal Of Experimental Ophthalmology
基金 国家自然科学基金项目(81970768) 北京大学第三医院眼科开放课题基金项目(BKLOF-BYSY2021-02)。
关键词 角膜移植 眼部感染 病毒性 聚合酶链式反应 植片失功 角膜内皮炎 Goldmann-Witmer系数 Corneal transplantation Eye infections,viral Polymerase chain reaction Graft failure Corneal endotheliitis Goldmann-Witmer coefficient
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