荧光定量PCR快速检测在病毒性角膜炎诊疗中的应用

目的 为病毒性角膜炎的患者进行疱疹病毒病原学快速检测，根据检测结果指导临床治疗。方法 选取未进行抗病毒药物治疗的患者，根据病情分别取其泪液或病变区角膜上皮细胞，用荧光定量PCR方法对单纯疱疹病毒（herpessimplex virus,HSV）1型、HSV2、水痘-带状疱疹病毒（varicella zoster virus,VZV）、人巨细胞病毒（human cytomegalovirus,HCMV）和EB病毒（EB virus,EBV）的核酸进行定量检测。结果 90例病毒性角膜炎患者中有54例检测出疱疹病毒核酸，检出率为60.0%；发病后首次就诊的病毒性角膜炎患者中疱疹病毒核酸的检出率为94.7%，明显高于多次发作后就诊的患者（检出率为34.6%），2者差异比较具有统计学意义（P<0.001）。发病后首次就诊的病毒性角膜炎患者感染的主要病毒为HSV1、EBV和HSV2，多次发作后就诊的患者感染的病毒主要为HSV1和HCMV。5种疱疹病毒中HSV占病毒检出数的74.0%，上皮型角膜炎中疱疹病毒核酸的检出率为30.6%，基质型角膜炎中疱疹病毒核酸的检出率为95.1%，基质型角膜炎中疱疹病毒检出率高于上皮型角膜炎(P<0.001）。上皮型角膜炎中HSV1病毒的检出率和平均拷贝数明显高于其他病毒；基质型角膜炎中HSV1、HSV2病毒的拷贝数明显高于其他疱疹病毒拷贝数。结论 病毒性角膜炎患者应尽早进行核酸检测以明确疱疹病毒类型及拷贝数，检测结果为进一步的抗病毒治疗和疗效判断提供科学依据。

Application of rapid detection of fluorescent quantitative PCR in diagnosis and treatment of viral keratitis

Objective To detect of herpes virus pathogens rapidly for patients with viral keratitis. Methods According to the severity of keratopathy, corneal epithelial cells from lesion area or tear were selected before drug treatment. The nucleic acids of herpes simplex virus-1(HSV1), HSV2, varicella zoster virus (VZV), human cytomegalovirus(HCMV) and EB virus(EBV) were quantitatively detected by fluorescence quantitative PCR. Results Five viral nucleic acids were detected in 54 out of 90 patients with viral keratitis, the detection rate was 60.0%. The detection rate of five viral nucleic acids was 94.7% in patients with viral keratitis treated for the first time after the onset of viral keratitis, which was significantly higher than that of patients treated after multiple attacks (the detection rate was 34.6%). There was a significant difference between two groups (P<0.001). HSV1, EBV and HSV2 were the main viruses in viral keratopathy patients who firstly visited after onset. HSV1 and HCMV were the main viruses in patients who visited after multiple outbreaks. HSV accounted for 74.0% of the five viruses detected, the detection rate of five viral nucleic acids in epithelial keratitis was 30.6%, the detection rate in stromal keratitis was 95.1%, and the detection rate in stromal keratitis was higher than that in epithelial keratitis (P<0.001). The detection rate and average copy number of HSV1 virus in epithelial keratitis were significantly higher than those of other viruses, and the copy number of HSV1 and HSV2 virus in stromal keratitis was significantly higher than that of other viruses. Conclusion Patients with viral keratitis should undergo nucleic acid testing as soon as possible to determine the type and number of copies of the virus. This result provide a scientific basis for further antiviral treatment and efficacy evaluation.