Clinical Metagenomic Next-Generation Sequencing for Diagnosis of Secondary Glaucoma in Patients With Cytomegalovirus-Induced Corneal Endotheliitis

Abstract

Glaucoma is the second leading cause of blindness globally. Growing scientific evidence indicated that inflammation of the trabecular meshwork induced by corneal endotheliitis could lead to secondary glaucoma. Cytomegalovirus (CMV) has been identified as the most common herpes virus in corneal endotheliitis patients. Early detection is critical in preventing endothelial cell loss, and patient management should vary based on different pathological factors. However, routine culture and real-time polymerase chain reaction (qPCR) have difficult in distinguishing whether CMV, Varicella Zoster Virus (VZV) or Herpes Simplex Virus (HSV) causes endothiliitis. This may result in inappropriate treatment, which may prolong or aggravate the status of disease. We compared the sensitivity and specificity of qPCR and Metagenomic Next-Generation Sequencing (mNGS) in the aqueous humor of patients with suspected CMV endotheliitis in this study. Our results showed that four out of 11 (36.4%) of our patients were positive for CMV by qPCR, whereas mNGS had a 100% detection rate of CMV. Our findings implied that mNGS could be a useful diagnostic tool for CMV-induced endotheliitis.

Keywords: cytomegalovirus; herpes simplex virus; metagenomic next-generation sequencing; secondary glaucoma; varicella-zoster virus.

FIGURE 1

The proposed procedure utilizing qPCR and mNGS analysis on the aqueous humor of CMV-infected patients.

FIGURE 2

Detection of the Human Cytomegalovirus (CMV) by metagenomic Next-generation sequencing (mNGS) on the aqueous humor from 11 glaucoma patients. The total DNA from aqueous humor samples from the eyes of the patient with suspected CMV infection was sequenced by BGISEQ-50/MGISEQ-2000 platform. The reads were mapped against human reference genome (hg19) using Burrows-Wheeler Alignment. From different samples, a different number of reads matching to CMV were recovered. The 3 number of the reads were considered as the “CMV-positive.” Genome coverage of detected CMV sequences and the relative abundance of detected viruses in the 11 patients (A–K).