Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Knowledge E
2021-01-01
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| Series: | Journal of Ophthalmic & Vision Research |
| Subjects: | |
| Online Access: | https://doi.org/10.18502/jovr.v16i1.8243 |
| _version_ | 1797990947068313600 |
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| author | Ahmed Nishat H Satpathy Gita Rohan Chawla Radhika Tandon |
| author_facet | Ahmed Nishat H Satpathy Gita Rohan Chawla Radhika Tandon |
| author_sort | Ahmed Nishat H |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T08:44:35Z |
| format | Article |
| id | doaj.art-b400a8e8a6a34914a1e1c5d95211744f |
| institution | Directory Open Access Journal |
| issn | 2008-322X |
| language | English |
| last_indexed | 2024-04-11T08:44:35Z |
| publishDate | 2021-01-01 |
| publisher | Knowledge E |
| record_format | Article |
| series | Journal of Ophthalmic & Vision Research |
| spelling | doaj.art-b400a8e8a6a34914a1e1c5d95211744f2022-12-22T04:34:00ZengKnowledge EJournal of Ophthalmic & Vision Research2008-322X2021-01-011631110.18502/jovr.v16i1.8243jovr.v16i1.8243Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral InfectionsAhmed Nishat H0Satpathy Gita1Rohan Chawla2Radhika Tandon3 Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, India Ophthalmology, Dr. R P Centre, All India Institute of Medical Sciences, New Delhi, IndiaPurpose: 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.https://doi.org/10.18502/jovr.v16i1.8243adenoviruscytomegalovirusherpes simplex virusmultiplex polymerase chain reactionvaricella-zoster virus |
| spellingShingle | Ahmed Nishat H Satpathy Gita Rohan Chawla Radhika Tandon 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 & Vision Research adenovirus cytomegalovirus herpes simplex virus multiplex polymerase chain reaction varicella-zoster virus |
| title | Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections |
| title_full | Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections |
| title_fullStr | Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections |
| title_full_unstemmed | Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections |
| title_short | Multiplex PCR for Detection of Herpes Simplex Viruses Type-1 and Type-2, Cytomegalovirus, Varicella-zoster Virus, and Adenovirus in Ocular Viral Infections |
| title_sort | multiplex pcr for detection of herpes simplex viruses type 1 and type 2 cytomegalovirus varicella zoster virus and adenovirus in ocular viral infections |
| topic | adenovirus cytomegalovirus herpes simplex virus multiplex polymerase chain reaction varicella-zoster virus |
| url | https://doi.org/10.18502/jovr.v16i1.8243 |
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