Indian Journal of Ophthalmology

ORIGINAL ARTICLE
Year
: 2019  |  Volume : 67  |  Issue : 7  |  Page : 1040--1046

Evaluation of multiplex real-time polymerase chain reaction for the detection of herpes simplex virus-1 and 2 and varicella-zoster virus in corneal cells from normal subjects and patients with keratitis in India


Sai Jeevana Madhuri Guda1, Bhavani Sontam1, Bhupesh Bagga2, Konduri Ranjith1, Savitri Sharma1, Joveeta Joseph1 
1 Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
2 Tej Kohli Cornea Institute, L. V. Prasad Eye Institute, Kallam Anji Reddy Campus, Hyderabad, Telangana, India

Correspondence Address:
Dr. Joveeta Joseph
Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L. V. Prasad Eye Institute, Hyderabad, Telangana - 500 034
India

Purpose: To determine the presence of herpes simplex virus and varicella zoster virus (HSV 1 and 2, VZV) in the cornea of normal subjects by multiplex real time quantitative (qPCR) assay and evaluate its utility in the diagnosis of viral keratitis. Methods: Corneal epithelial cells from 33 eyes of 22 patients undergoing photorefractive keratectomy surgery (controls) and 50 corneal scrapings from 50 patients with suspected HSV keratitis were analyzed for the presence of HSV1 by conventional PCR and for presence of HSV1 and 2 and/or VZV by multiplex real-time PCR. Corneal scrapings of patients were also tested for HSV1 antigen by immunofluorescence assay (IFA). The results were compared and clinical records reviewed. Results: HSV1 and VZV DNA were detected in 8/33 controls (mean-14.3 ± 7.96, range: 3-29.1 copies/mL) and 2/33 controls (mean-10.7 ± 10.9, range 3-18.5 copies/ml) respectively. HSV2 was not detected in any of the controls. Copy numbers above the mean + 1SD of controls were considered significant for viral load in patient samples. Significantly higher number of corneal scrapings (39/50, 78%) from patients were positive for HSV1 (1.2 × 106 copies/mL ± 3.7 × 106 copies/mL) by real time qPCR compared to IFA (11/48, 23%, P value 0.0001) and conventional PCR (20/50, 40%, P value 0.0002). Double infection with HSV-1 (1.5 × 107 copies/ml) and HSV-2 (3.57 × 104 copies/ml) in one case and VZV infection (1.03 × 102 copies/ml) in another was also detected by the multiplex real-time PCR. Conclusion: Multiplex real-time PCR reliably detects HSV1 and 2 and VZV DNA and is ideal for the diagnosis of HSV and VZV keratitis in an ocular microbiology laboratory.


How to cite this article:
Guda SJ, Sontam B, Bagga B, Ranjith K, Sharma S, Joseph J. Evaluation of multiplex real-time polymerase chain reaction for the detection of herpes simplex virus-1 and 2 and varicella-zoster virus in corneal cells from normal subjects and patients with keratitis in India.Indian J Ophthalmol 2019;67:1040-1046


How to cite this URL:
Guda SJ, Sontam B, Bagga B, Ranjith K, Sharma S, Joseph J. Evaluation of multiplex real-time polymerase chain reaction for the detection of herpes simplex virus-1 and 2 and varicella-zoster virus in corneal cells from normal subjects and patients with keratitis in India. Indian J Ophthalmol [serial online] 2019 [cited 2020 Aug 14 ];67:1040-1046
Available from: http://www.ijo.in/article.asp?issn=0301-4738;year=2019;volume=67;issue=7;spage=1040;epage=1046;aulast=Guda;type=0