目的研究疱疹病毒DNA在疱疹病毒阳性角膜移植患者不同角膜层次中的分布，比较角膜和房水样本在该人群中的病毒DNA检测效率。
方法采用诊断试验研究方法，通过临床病历系统收集2015年5月至2021年8月于北京大学第三医院眼科中心行角膜移植且术中角膜和/或房水疱疹病毒DNA阳性患者。分析疱疹病毒DNA阳性患者的一般资料和分布层次。计算疱疹病毒在角膜和房水中的检出率，通过受试者工作特征曲线分析角膜和房水病毒DNA检测灵敏度，记录曲线下面积(AUC)和95%置信区间(CI)。
结果角膜移植术中疱疹病毒DNA阳性患者共166例166眼，其中巨细胞病毒(CMV)75眼(占45.2%)，单纯疱疹病毒1型(HSV-1)34眼(占20.5%)，水痘－带状疱疹病毒(VZV)30眼(占18.1%)，EB病毒(EBV)27眼(占16.3%)。CMV和VZV DNA在角膜内皮层分布者分别为47眼(占62.7%)和26眼(占86.7%)，明显多于基质层的28眼(占37.3%)和4眼(占13.3%)，差异均有统计学意义(χ2＝4.813、16.133，均P<0.05)。HSV-1和EBV DNA在角膜内皮层分布者分别为8眼(占23.5%)和5眼(占18.5%)，分别少于基质层的26眼(占76.5%)和22眼(占81.5%)，差异均有统计学意义(χ2＝9.529、10.704，均P<0.001)。疱疹病毒DNA阳性样本的检测灵敏度结果显示，角膜样本的病毒PCR检测总体灵敏度为71.6%，高于房水的54.1%，其中，CMV和VZV DNA阳性的角膜样本检测灵敏度分别为64.3%(AUC＝0.821，95% CI：0.705~0.938)和35.7%(AUC＝0.679，95% CI：0.475~0.882)，分别低于房水样本的71.4%(AUC＝0.875，95% CI：0.750~0.964)和85.7%(AUC＝0.929，95% CI：0.816~1.000)。HSV-1和EBV DNA阳性的角膜样本检测灵敏度分别为100%(AUC＝1.000，95% CI：1.000~1.000)和92.3%(AUC＝0.962，95% CI：0.875~1.000)，明显高于房水样本的27.8%(AUC＝0.639，95% CI：0.455~0.823)和23.1%(AUC＝0.615，95% CI：0.395~0.835)。
结论在疱疹病毒DNA阳性的角膜移植患者中，CMV DNA的检出率最高，CMV和VZV DNA多在角膜内皮层被检出，HSV-1和EBV DNA多在角膜基质层被检出。相较房水样本，角膜组织病毒DNA的检测灵敏度更高。
ObjectiveTo investigate the distribution of herpesvirus DNA in the corneal layers of herpesvirus-positive corneal transplantation patients and to compare the efficiency of viral DNA detection in corneal and aqueous humor samples in these patients.
MethodsA diagnostic test study was conducted.Data from patients, who underwent keratoplasty in Peking University Third Hospital from May 2015 to August 2021 and tested positive for herpesvirus in corneal tissue and/or aqueous humor samples obtained during surgery, were collected through the clinical medical record system.The demographic data and virus distribution layers of these patients were analyzed.The rate of herpesvirus detection in different samples was analyzed.The sensitivity of different samples for viral DNA detection was analyzed by receiver operating characteristic curves, and area under the curve (AUC) and 95% confidence interval (CI) were recorded.This study adhered to the Declaration of Helsinki and the study protocol was approved by the Ethics Committee of Peking University Third Hospital (No.M2021283).Written informed consent was obtained from each patient before entering the cohort.
ResultsA total of 166 herpesvirus-positive patients (166 eyes) were collected.Of the 166 eyes, 75 eyes (45.2%) were positive for cytomegalovirus (CMV), 34 eyes (20.5%) for herpes simplex virus 1 (HSV-1), 30 eyes (18.1%) for varicella zoster virus (VZV), and 27 eyes (16.3%) for Epstein-Barr virus (EBV).CMV DNA and VZV DNA were detected in the endothelial layers of 47 eyes (62.7%) and 26 eyes (86.7%), respectively, which were significantly higher than the 28 eyes (37.3%) and 4 eyes (13.3%) with virus located in stromal layers (χ2=4.813, 16.133; both at P<0.05).HSV-1 DNA and EBV DNA were detected in the endothelial layer of 8 eyes (23.5%) and 5 eyes (18.5%), respectively, which were less than the 26 eyes (76.5%) and 22 eyes (81.5%) with virus located in stromal layers, and the differences were statistically significant (χ2=9.529, 10.704; both at P<0.001).The sensitivity of corneal samples for herpesvirus DNA positivity was 71.6%, which was higher than 54.1% of aqueous humor.The detection sensitivities of corneal samples for CMV DNA and VZV DNA positivity were 64.3%(AUC=0.821, 95% CI: 0.705-0.938) and 35.7%(AUC=0.679, 95% CI: 0.475-0.882), respectively, which were lower than 71.4%(AUC=0.875, 95% CI: 0.750-0.964) and 85.7%(AUC=0.929, 95% CI: 0.816-1.000) of aqueous humor samples.The detection sensitivities of corneal samples for HSV-1 DNA and EBV DNA were 100%(AUC=1.000, 95% CI: 1.000-1.000) and 92.3%(AUC=0.962, 95% CI: 0.875-1.000), respectively, which were higher than 27.8%(AUC=0.639, 95% CI: 0.455-0.823) and 23.1%(AUC=0.615, 95% CI: 0.395-0.835) of aqueous humor samples.
ConclusionsThe detection rate of CMV DNA is highest among herpesvirus-positive keratoplasty patients.CMV DNA and VZV DNA are primarily located in the corneal endothelial layers, while HSV-1 DNA and EBV DNA are more predominant in the corneal stromal layer.The sensitivity of virus DNA detection is higher in the cornea than in aqueous humor.
臧云晓,彭荣梅,肖格格,等. 房水与角膜样本对疱疹病毒阳性角膜移植后不同层次病毒DNA检测效率比较[J]. 中华实验眼科杂志,2023,41(10)：992-997.
DOI:10.3760/cma.j.cn115989-20220115-00018版权归中华医学会所有。
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臧云晓：直接参与选题、酝酿和设计试验、实施研究、采集数据、分析/解释数据、起草及修改文章；彭荣梅：设计试验、分析/解释数据，对文章内容审阅和修改；肖格格：采集数据、起草文章、对文章内容审阅和修改；何林辉：设计试验、实施研究、采集数据、起草文章；洪晶：采集数据、对文章内容审阅和修改及定稿
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