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Nishat H A, Satpathy G, Chawla R, Tandon R. Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections. JOVR [Internet]. 2021Jan.20 [cited 2024May5];16(1):3–11. Available from: https://knepublishing.com/index.php/JOVR/article/view/8243

https://doi.org/10.18502/jovr.v16i1.8243

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authors

  • Ahmed Nishat H

    Ahmed Nishat H

    Ocular Microbiology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India
    https://orcid.org/0000-0002-1683-4275
  • Gita Satpathy

    Gita Satpathy

    Ocular Microbiology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India
  • Rohan Chawla

    Rohan Chawla

    Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India
  • Radhika Tandon

    Radhika Tandon

    Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India

Published Date

  • Jan 20, 2021
This journal is publishing all the articles under Creative Commons Attribution License CC BY 4.0.

Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections

Journal of Ophthalmic and Vision Research (JOVR) / January–March 2021, Volume 16, Issue 1 / Pages 3–11

Abstract

Purpose: Most common viruses causing ocular infections are Herpes Simplex Viruses (HSV) type 1 and type 2, Cytomegalovirus (CMV), Varicella-zoster Virus (VZV), and few strains of Adenovirus. Diagnosis of these infections through clinical manifestations and using conventional methods has a number of limitations. The purpose of this study was to develop a multiplex Polymerase Chain Reaction (PCR) for simultaneous detection of all pathogenic viruses from ocular infections.


Methods: Ten uniplex PCRs were standardized, two each for HSV type 1 (HSV-1) and type 2 (HSV-2), CMV, VZV, and Adenovirus. Various multiplexing combinations of above PCRs were put to finalize targets and reaction conditions enabling diagnosis of all in a single reaction. The uniplex and multiplex PCRs were run for known positive and negative controls, and samples from clinically suspected patients and healthy controls.


Results: Out of the 170 samples from suspected ocular infections, 24.7% were positive by uniplex PCR and 22.9% were correctly identified by multiplex PCR. None of the samples negative by uniplex PCRs was positive by the multiplex PCR. The sensitivity and specificity of multiplex PCR compared to the commonly used uniplex PCRs as gold standard was 92.86% and 100%, respectively. The prevalence of different viral pathogens was 13.5% for HSV-1, followed by 5.9% for Adenovirus, 2.4% for VZV, 1.8% for HSV-2, and 1.2% for CMV.


Conclusion: The establishment of multiplex PCR has found immediate application in diagnosing ocular viral pathogens in a single reaction, thus saving time, manpower, and resources by fivefold.

Keywords:

Adenovirus, Cytomegalovirus, Herpes Simplex Virus, Multiplex Polymerase Chain Reaction, Varicella-zoster Virus

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