Comparison Of PCR Detection Results Of Tears,Aqueous Humor And Anterior Capsule In The Quiescent Phase Of Herpetic Ocular Diseases

Objective:To compare the quantification of herpes simplex virus(HSV)DNA in the tear fluid,aqueous humor and anterior capsule in the quiescent phase of herpetic stromal keratitis,endotheliitis and anterior uveitis using quantitative real-time Polymerase Chain Reaction(qPCR).Methods:61 eyes with clinically diagnosed herpetic ocular diseases who were treated at our hospital between September 2016 and April 2019 were enrolled in this study.These patients were divided into three groups:22 eyes with herpetic stromal keratitis,18 eyes with endotheliitis and 21 eyes with anterior uveitis.When the active intraocular inflammation of these patients was resolved,and these patients had decreased visual acuity due to the cataract formation,we performed the cataract surgery and collected the clinical data and samples included tear fluid,aqueous humor and anterior capsule of each patient.The control group consisted of 60 eyes from 60 candidates for cataract surgery who had no herpetic ocular diseases features in either eye,we collected the same samples during the surgery.Viral DNA was extracted from samples using PureLinkTM Viral RNA/DNA mini kit and amplified by qPCR.The quantification of herpes simplex virus(HSV)DNA and clinal data between four groups were compared using variance analysis and Chi-square test.Kendall's W test was used to analyze the consistency of the positive rate of HSV DNA in three different samples.The relationship between the positive rate of HSV DNA and the sample type,disease type,and duration of the quiescent phase was analyzed by a two-class logistic regression model.The statistical significance was set to 0.05.Results:There was no statistical differences in gender,age,involved eyes and duration of the quiescent phase between four groups.There was no statistical differences in the positive rate of HSV DNA in tears between four groups.The positive rate of HSV DNA in aqueous humor and anterior capsule of endotheliitis group(38.9%and 38.1%respectively)and anterior uveitis group(38.9%and 38.1%respectively)were significantly higher than stromal keratitis group(0%and 3.3%respectively)and control group(4.5%and 11.7%respectively;P=0.000 and 0.002 respectively).The positive rate of HSV DNA in all three kinds of sample in patients with duration of the quiescent phase longer than 2 months were lower than patients with duration of the quiescent phase equal or shorter than 2 months,but only the differences in aqueous humor were statistically significant(6.2%vs.34.5%,P=0.014).There were no significant consistency in the positive rates of HSV DNA in the three kinds of sample.The positive rate of HSV DNA in anterior capsule was much higher than that in tear fluid and aqueous humor.The positive rate of HSV DNA in all three kinds of sample of involved eyes were higher than those in contralateral eyes,but only the differences in aqueous humor were statistically significant(41.7%vs.0%,P=0.019).There was no statistical differences in the positive rate of HSV DNA in all three kinds of samples between the contralateral eyes and control group.Conclusion:In the quiescent phase of herpetic ocular diseases,the detection rate of HSV DNA in patients with anterior uveitis and endotheliitis was much higher than those in stromal keratitis.The positive rate of HSV DNA in anterior capsule was much higher than that in tear fluid and aqueous humor.The detection rate of HSV DNA is higher in patients with shorter quiescent phase.