聚合酶链反应检测角膜内皮炎泪液及房水中单纯疱疹病毒DNA

目的 :探讨聚合酶链反应 (PCR)技术对角膜内皮炎的病原学诊断价值。方法 :用PCR技术对角膜内皮炎房水及泪液中的单纯疱疹病毒Ⅰ型DNA进行扩增 ,并用琼脂糖凝胶电泳对PCR产物进行检测 ;同时以老年性白内障房水及泪液为对照组。结果 :16例角膜内皮炎房水中 11例阳性 ,阳性率为 68 75 % ,2 0例对照组房水中无 1例阳性 ,二者有显著性差异 (P <0 0 5 ) ;16例角膜内皮炎泪液中 3例阳性 ,阳性率为 18 75 % ,2 0例对照组泪液中 1例阳性 ,阳性率为 5 % ,二者无显著性差异 (P >0 0 5 ) ;角膜内皮炎房水检则阳性率 (68 75 % )与泪液检测阳性率 (18 75 % )有显著性差异 (P <0 0 5 )。结论 :用PCR技术检测角膜内皮炎房水中单疱病毒DNA可以对角膜内皮炎做出病原学诊断 ,并可进一步指导临床治疗。     

间接免疫荧光技术检测角膜内皮炎房水及小梁组织中的单纯疱疹病毒

5目的 :探讨间接免疫荧光技术 (IIF)对角膜内皮炎的病原学诊断价值 ,并以此揭示病因、指导临床治疗。方法 :用IIF对角膜内皮炎的房水及小梁中的单纯疱疹病毒 (HSV )进行检测 ,以老年性白内障和青光眼小梁组织作对照。结果 :实验组 13例角膜内皮炎患者的房水中有 4例呈阳性发现 ,其阳性率为 3 0 77% ;而对照组Ⅰ 2 0例老年性白内障患者的房水中无一呈阳性发现 ,二者之间存在显著性差异 (P <0 0 5 )。实验组 1例角膜内皮炎患者的小梁组织中呈阳性发现 ,而对照组Ⅱ 10例青光眼患者的小梁组织中无一阳性发现。结论 :应用IIF可检测角膜内皮炎患者急性期房水和小梁组织中的HSV ,说明角膜内皮炎的发病同HSV感染有关 ,同时可做出实验室病原学诊断 ,并进一步用于指导临床治疗。     

聚合酶链反应检测角膜内皮炎房水中单纯疱疹病毒Ⅰ型DNA及带状疱疹病毒DNA

目的 评价聚合酶链反应技术（PCR）对角膜内皮炎的诊断价值，并以此解释病因，指导临床治疗。方法对临床诊断为角膜内皮炎的患者12例（12眼）及临床诊断为年龄相关性白内障的患者15例（15眼）分别用PCR方法进行房水中单纯疱疹病毒Ⅰ型（HSV-Ⅰ）DNA及带状疱疹病毒（VZV）DNA的检测，结果采用分类变量资料两样本率比较的四格表确切概率比较。结果共扩增角膜内皮炎患者12例（12眼），HSV-Ⅰ阳性者5例（5眼），阳性率为41．67％，共扩增对照组15例（15眼），HSV-Ⅰ阳性者0例，阳性率为0％，二者差异有显著性。结论PCR技术检测角膜内皮炎患者房水中的HSV-Ⅰ，可在分子水平上建立一种快速准确的诊断方法，为临床治疗提供可靠的依据。     

多聚酶链反应和免疫荧光法对单纯疱疹病毒性角膜炎诊断的比较研究

目的 :探讨单纯疱疹病毒性角膜炎的快速实验室诊断的方法和结果。方法 :对临床诊为单疱角膜炎的 3 0例 3 0眼的泪液及角膜上皮行PCR法检测HSV -1DNA ,并对角膜上皮细胞中的HSV -1抗原行IFA检测 ,同时以 10例健康眼作为正常对照 ,并作统计学分析。结果 :PCR阳性检出率 80 % (2 4/ 3 0 ) ,IFA检出率 40 % (12 / 3 0 ) ,对比P <0 0 0 5。泪液PCR阳性率 66 7% (2 0 / 3 0 )同角膜上皮PCR阳性率 60 % (18/ 3 0 )对比P >0 5 ,无显著差异。结论 :PCR法比IFA法敏感 ,可作为单疱角膜炎的病原学快速诊断 ,泪液可作为首选标本。     

荧光定量PCR技术检测HSK房水中HSV-Ⅰ型DNA

目的探讨荧光定量聚合酶链反应（FQ-PCR）技术对I型单纯疱疹病毒性角膜炎（HSK）的病原学诊断价值。方法应用FQ-PCR技术对HSK患眼房水中的I型单纯疱疹病毒（HSV-I）DNA进行扩增,并对PCR产物进行定量检测,同时以老年性白内障患者房水作为对照,并作统计学分析。结果 16例（16只眼）HSK患眼房水中有5例阳性,阳性率31.25%,20例（20只眼）单纯老年性白内障患者房水无1例阳性,阳性率0%,差异有统计学意义（P〈0.05）。结论 FQ-PCR法可以作为HSK的病原学快速诊断的实验室方法之一。     

荧光定量PCR技术检测HSK房水中HSV-I型DNA

目的 探讨荧光定量聚合酶链反应(FQ-PCR)技术对I型单纯疱疹病毒性角膜炎(HSK)的病原学诊断价值.方法 应用FQ-PCR技术对HSK患眼房水中的I型单纯疱疹病毒(HSV-I)DNA进行扩增,并对PCR产物进行定量检测,同时以老年性白内障患者房水作为对照,并作统计学分析.结果 16例(16只眼)HSK患眼房水中有5例阳性,阳性率31.25%,20例(20只眼)单纯老年性白内障患者房水无1例阳性,阳性率0%,差异有统计学意义(P<0.05).结论 FQ-PCR法可以作为HSK的病原学快速诊断的实验室方法 之一.     

内眼手术患者血液和房水梅毒抗体检测的对比研究

目的 探讨内眼手术患者中发生梅毒的阳性率,并通过梅毒患者房水中特异性和非特异性抗体的检测,寻找梅毒是否通过房水传播的理论依据.方法 对2004年以来准备接受内眼手术的4075例患者进行常规的传染四项检查,包括乙肝表面抗原(HBsAg),丙型肝炎抗体(HCV抗体),梅毒抗体(TP抗体),艾滋病抗体(HIV抗体)四项指标,并统计梅毒发生的阳性率;应用特异性抗体试验-梅毒螺旋体血球凝结试验(TPHA)和非特异性抗体试验-快速血浆反应素环状卡片试验(RPR)检测梅毒患者血清及房水中的抗体.结果 4075例内眼手术患者有58例患者血液中存在梅毒抗体,阳性率1.42%.58例梅毒患者血液中TPHA阳性检出率为100%(58例),房水中为32.76%(19例)两者比较差异有统计学意义(P＜0.05);而同组患者血液中RPR阳性检出率为51.72%(30例).房水中为8.62%(5例)两者比较差异有统计学意义(P＜0.05).结论 近年来梅毒感染阳性率相对稳定并且梅毒可能不通过房水传播.     

病毒性前葡萄膜炎患者房水病毒类型检测分析

目的 分析临床诊断为病毒感染性前葡萄膜炎患者的房水病毒检测结果。设计 回顾性病例系列。研究对象 临床诊断为病毒感染性前葡萄膜炎患者25例（25眼）。方法 记录患者全身病毒感染病史,对患者进行血清免疫学检查（HLA-B27、RF等）、常规眼科检查（包括视力、眼压、裂隙灯、间接检眼镜检查等）,通过前房穿刺获取房水,应用RT-PCR技术检测房水病毒。通过 Logistic回归分析评估临床表现与PCR检测病毒阳性的关系。主要指标 房水RT-PCR病毒检测结果。结果12例（48%）患者有病毒感染病史。25例（25眼）患者中14例（56%）房水RT-PCR病毒检测阳性。其中水痘-带状疱疹病毒9例（36%）,患者既往均有水痘或带状疱疹病史;单纯疱疹病毒4例（16%);巨细胞病毒1例（4%）。PCR检测病毒阳性的患者更易出现角膜水肿（19眼,76%）（OR=2.006,P=0.043）、眼压升高（19眼,76%,平均（32.3±19.7）mmHg）（OR=1.905,P=0.013）、色素性KP（9眼,36%）（OR=1.31,P=0.014）或细小灰白KP（8眼,32%）（OR=1.41,P=0.014）。结论 RT-PCR房水病毒检测对临床辅助诊断病毒前葡萄膜炎有一定价值。病毒性前葡萄膜炎最常见的致病病毒是水痘-带状疱疹病毒。主要临床表现为角膜水肿、眼压高、色素性KP或细小灰白KP。（眼科,2021, 30： 421-424）     

先天性青光眼患儿房水及血清疱疹病毒抗体检测分析

目的研究原发性先天性青光眼(PCG)患儿房水及血清中的疱疹病毒感染情况,并比较分析病毒抗体阳性和阴性PCG患儿临床特点的差异。方法抽取13例PCG患儿首次手术眼13眼的房水及血清,采用酶联免疫吸附试验(ELISA)检测房水单纯疱疹病毒(HSV)、水痘-带状疱疹病毒(VZV)、巨细胞病毒(CMV)Ig G抗体,血清中HSV、VZV、CMV Ig G和Ig M抗体,并采用聚合酶链反应(PCR)验证房水中病毒情况。抽取13例老年性白内障患者的房水作为房水对照组;抽取50例同年龄健康儿童外周血作为血清对照组。对PCG疱疹病毒抗体阳性组和阴性组间年龄、性别、眼别、最高眼压、杯盘比、角膜直径、房角结构、Haab纹等临床特点进行比较分析。结果 13例PCG患儿中9例(69.2%)血清疱疹病毒抗体检测阳性,对照组50例儿童无血清疱疹病毒抗体检测阳性病例;在所有13例PCG患儿中,房水病毒抗体阳性1例(CMV-Ig G),另1例CMV PCR阳性,此2例患者血清疱疹病毒Ig G抗体检测均为阳性,其余房水样本及对照组房水病毒检测均为阴性。PCG患儿中,起病最高眼压、角膜直径、杯盘比和Haab纹所占比例在病毒阳性组和阴性组间差异均无统计学意义。结论超过一半(69.2%)的PCG患者存在血清疱疹病毒抗体阳性。病毒阳性组与阴性组的起病最高眼压、角膜直径、杯盘比及Haab纹占比基本相似。     

Optisol中期保存液房水置换对角膜内皮细胞的保护作用

目的:研究Optisol中期保存液房水置换对角膜内皮细胞的保护作用。方法:采用4℃湿房保存方法保存离体兔眼球,设立房水对照组、Optisol中期保存液房水置换组,每组10只眼球。于低温保存开始前、保存后24,48及72h检测角膜厚度、角膜内皮细胞密度及形态,保存后72h取下角膜行胎盘蓝-茜素红染色,检测角膜内皮细胞存活率。结果:4℃湿房保存开始前两组平均角膜厚度、角膜内皮细胞密度及六角形细胞比率差异没有统计学意义(P>0.05)。保存后24h两组平均角膜厚度、角膜内皮细胞密度及六角形细胞比率差异没有统计学意义(P>0.05);保存后48h及72h两组平均角膜厚度、角膜内皮细胞密度及六角形细胞比率差异有统计学意义(P<0.05)。保存后72h房水对照组角膜内皮细胞存活率为(55±5.81)%,Optisol中期保存液房水置换组角膜内皮细胞存活率为(87.16±3.64)%,两组间比较有统计学意义(P<0.05)。结论:Optisol中期保存液房水置换对角膜内皮细胞活性有保护作用,可以提高湿房保存法的效率,延长有效保存时间至72h。     

Cytomegalovirus in aqueous humor from an eye with corneal endotheliitis

PURPOSE: To report cytomegalovirus (CMV) DNA in aqueous humor from a patient with unilateral corneal endotheliitis. DESIGN: Case report. METHODS: A 51-year-old man presented with unilateral corneal endotheliitis with linear keratic precipitates and coin-shaped lesions. Tear and aqueous humor samples were subjected to polymerase chain reaction to look for DNA from herpes simplex virus (HSV), varicella zoster virus (VZV), and CMV. RESULTS: Aqueous humor from the diseased eye contained DNA from CMV but not HSV or VZV. Its specificity was confirmed by Southern blot tests. Intravenous ganciclovir treatment resulted in the localization of his corneal edema and the reduction in keratic precipitates. There was severe destruction of corneal endothelial cells. CMV DNA was not detected in tears or control samples. CONCLUSIONS: In this healthy man with corneal endotheliitis, we detected CMV DNA in aqueous humor from the affected eye, but not HSV or VZV. This suggests that CMV may cause corneal endotheliitis in patients without immunodeficiency.     

Corneal endotheliitis and idiopathic sudden sensorineural hearing loss

PURPOSE: To report a case with corneal endotheliitis and idiopathic sudden sensorineural hearing loss, in which herpes simplex virus type 1 DNA was demonstrated in the trabeculum and the aqueous humor by polymerase chain reaction. DESIGN: Interventional case report. METHODS: A 60-year-old man presented with corneal stromal edema in the right eye and sudden bilateral sensorineural hearing loss. The trabeculum excised during trabeculectomy and the aqueous humor were examined for the presence of herpes simplex virus type 1 DNA by polymerase chain reaction. RESULTS: Polymerase chain reaction demonstrated herpes simplex virus type 1 DNA in the aqueous humor and the trabeculum. CONCLUSION: Herpes simplex virus type 1 may cause corneal endotheliitis and idiopathic sudden sensorineural hearing loss simultaneously.     

DNA of cytomegalovirus detected by PCR in aqueous of patient with corneal endotheliitis after penetrating keratoplasty

PURPOSE: Corneal endotheliitis often leads to severe endothelial dysfunction and can be caused by herpes simplex virus (HSV), varicella zoster virus (VZV), and other viruses (eg, the mumps virus). We report a case of corneal endotheliitis caused by cytomegalovirus (CMV) that developed after a penetrating keratoplasty. METHODS: A complete ophthalmologic examination was performed on a patient with corneal endotheliitis that developed after a penetrating keratoplasty. To determine the cause of the endotheliitis, polymerase chain reaction (PCR) was used to amplify the DNA of HSV, VZV, and CMV in samples of the aqueous humor. RESULTS: Slit-lamp biomicroscopy showed a moderate stromal edema in the upper temporal part of the transplanted cornea along with keratic precipitates (KPs) arranged in a coin-shaped pattern. Repeated treatments with steroids and acyclovir were only temporarily successful. PCR detected the DNA of CMV in an aqueous sample, and the treatment was switched to topical and systemic application of ganciclovir. This resulted in the disappearance of the KPs and resolution of the stromal edema within 2 weeks. CONCLUSIONS: From the PCR results and the favorable response to ganciclovir, the corneal endotheliitis was most likely caused by cytomegalovirus in this case.     

Identification of cytomegalovirus and human herpesvirus-6 DNA in a patient with corneal endotheliitis

Purpose

To report the case of a patient with unilateral corneal endotheliitis in which both cytomegalovirus (CMV) and human herpesvirus-6 (HHV6) DNA was identified in the aqueous humor.

Case

A 67-year-old man with corneal endotheliitis OD was referred to us for decreased visual acuity. Local corneal stromal edema, pigmented keratic precipitates, a coin-shaped lesion and minimal anterior chamber reaction were observed by slit-lamp biomicroscopy. Cells with owl’s eye appearance in the endothelial cell layer were observed by in vivo laser confocal microscopy. The patient had rheumatoid arthritis, which was treated by oral prednisolone and intravenous abatacept. Polymerase chain reaction analysis of aqueous humor samples detected both CMV and HHV6 DNA, but not other HHVs. Treatment with topical ganciclovir and systemic valganciclovir resulted in a clear cornea.

Conclusions

A patient with corneal endotheliitis had both CMV and HHV6 DNA identified in the aqueous humor. Although both viruses were identified in this case, clinical manifestations resembled CMV corneal endotheliitis, and it was unclear whether HHV6 could affect the clinical course. Systemic abatacept and corticosteroid therapy might play a positive role in cases with both CMV and HHV6 DNA in this corneal endotheliitis.     

Progressive herpetic linear endotheliitis

PURPOSE: To report the clinical course of a rare case of bilateral herpetic linear endotheliitis. METHODS: A 70-year-old man presented with bilateral circumferential bullous edema with stromal edema progressing centrally in the left cornea and bilateral sensorineural hearing impairment simultaneously. Serum immunoglobulin G (IgG) and IgM antibodies against herpes simplex virus type 1 (HSV1) were tested for, and aqueous humor from both eyes was examined separately using polymerase chain reaction for the presence of HSV1 DNA. RESULTS: Serum antibody titers against HSV1 were positive. In the polymerase chain reaction, the aqueous humor showed HSV1 DNA in both eyes. Forty milligrams of prednisolone was given per day and 200 mg of oral acyclovir was given 4 times daily, but corneal edema progressed. After penetrating keratoplasty surgery in the left eye, recurrent herpetic endotheliitis also seemed to occur. CONCLUSIONS: HSV-1 may cause bilateral corneal linear endotheliitis and hearing impairment simultaneously. Linear endotheliitis should be regarded as a manifestation of HSV1 corneal infection. There is a poor prognosis, and severe corneal edema can result if aggressive treatment is not used.     

Experimental corneal endotheliitis in rabbit

PURPOSE: Corneal endotheliitis may cause permanent visual loss due to endothelial decompensation. The pathogenesis underlying this distinct clinical entity is not known. In the current study, a rabbit herpetic corneal endotheliitis model was made of induced anterior chamber-associated immune deviation (ACAID). METHODs: One group of rabbits received left-eye intracameral inoculation of UV-inactivated herpes simplex virus (HSV)-1 (strain McKrae). The second group received cell medium in the same manner as the first group. The third group subcutaneously received the same inoculum as the first group. Seven days later, all right eyes were intracamerally infected with 2.5 x 10(4) plaque-forming units of infectious HSV-1. Eyes were evaluated by slit lamp examination. Two weeks after infection, rabbits were killed, and right eyes were examined by immunohistochemical staining and electron microscopy. Aqueous humor was detected for HSV-1 DNA and antibody. RESULTS: Nonspecific inflammation occurred in the anterior segments of the eyes from the second and third groups. In contrast, at 14 days after infection, the first group of rabbits showed a specific pattern of inflammation that greatly resembled clinical features of corneal endotheliitis. Viral antigen was detected only in the endothelial layer. Electron microscopy revealed enlarged intercellular gaps and infiltration of inflammatory cells that are characteristic of endothelial defects. HSV-1 DNA was detected at a significantly higher number in the aqueous humor aspirates from endotheliitis rabbits. In addition, ACAID was shown to be induced in the rabbits with corneal endotheliitis. CONCLusIONS: HSV-1 infection can induce corneal endotheliitis and ACAID may play the pivotal role in this entity.     

糖尿病合并白内障患者血清及房水中铬元素含量测定及分析

目的:检测2型糖尿病合并老年性白内障患者血清及眼房水中铬元素的含量,并探讨其临床意义。方法:采用石墨炉原子吸收法测定患者血清及房水中铬的含量,试验组:糖尿病(2型)合并白内障患者19例,对照组:健康的老年性白内障患者21例。结果:试验组的血清及房水铬元素含量分别为(3.79±1.17)μg/L,(0.97±0.35)μg/L,明显低于对照组(4.50±0.92)μg/L,(1.43±0.68)μg/L,且两组比较差异均有统计学意义(t=-2.139,P<0.05;t=-2.653,P<0.05),两组血清及房水中铬元素含量无显著相关性(r=-0.142,P>0.05)。结论:铬元素在糖尿病合并老年性白内障患者的发生及发展过程中具有重要作用。