Generic selectors
Exact matches only
Search in title
Search in content
Search in posts
Search in pages
Filter by Categories
15th National Conference of the IAOMFP, Chennai, 2006
Abstracts from current literature
Acne in India: Guidelines for management - IAA Consensus Document
Art & Psychiatry
Association Activities
Association Notes
Award Article
Book Review
Brief Report
Case Analysis
Case Letter
Case Letters
Case Notes
Case Report
Case Reports
Clinical and Laboratory Investigations
Clinical Article
Clinical Studies
Clinical Study
Conference Oration
Conference Summary
Continuing Medical Education
Cosmetic Dermatology
Current Best Evidence
Current View
Derma Quest
Dermato Surgery
Dermatosurgery Specials
Dispensing Pearl
Do you know?
Drug Dialogues
Editor Speaks
Editorial Remarks
Editorial Report
Editorial Report - 2007
Editorial report for 2004-2005
Fourth All India Conference Programme
From Our Book Shelf
From the Desk of Chief Editor
Get Set for Net
Get set for the net
Guest Article
Guest Editorial
How I Manage?
IADVL Announcement
IADVL Announcements
IJDVL Awards
IJDVL Awards 2018
IJDVL Awards 2019
IJDVL Awards 2020
IJDVL International Awards 2018
Images in Clinical Practice
In Memorium
Inaugural Address
Knowledge From World Contemporaries
Leprosy Section
Letter in Response to Previous Publication
Letter to Editor
Letter to the Editor
Letter to the Editor - Case Letter
Letter to the Editor - Letter in Response to Published Article
Letter to the Editor - Observation Letter
Letter to the Editor - Study Letter
Letter to the Editor - Therapy Letter
Letter to the Editor: Articles in Response to Previously Published Articles
Letters in Response to Previous Publication
Letters to the Editor
Letters to the Editor - Letter in Response to Previously Published Articles
Letters to the Editor: Case Letters
Letters to the Editor: Letters in Response to Previously Published Articles
Medicolegal Window
Miscellaneous Letter
Net Case
Net case report
Net Image
Net Letter
Net Quiz
Net Study
New Preparations
News & Views
Observation Letter
Observation Letters
Original Article
Original Contributions
Pattern of Skin Diseases
Pediatric Dermatology
Pediatric Rounds
Presedential Address
Presidential Address
Presidents Remarks
Report of chief editor
Report of Hon : Treasurer IADVL
Report of Hon. General Secretary IADVL
Research Methdology
Research Methodology
Resident page
Resident's Page
Resident’s Page
Residents' Corner
Residents' Corner
Residents' Page
Review Article
Review Articles
Revision Corner
Self Assessment Programme
Seminar: Chronic Arsenicosis in India
Seminar: HIV Infection
Short Communication
Short Communications
Short Report
Special Article
Specialty Interface
Study Letter
Symposium - Contact Dermatitis
Symposium - Lasers
Symposium - Pediatric Dermatoses
Symposium - Psoriasis
Symposium - Vesicobullous Disorders
Symposium Aesthetic Surgery
Symposium Dermatopathology
Symposium-Hair Disorders
Symposium-Nails Part I
Symposium-Nails-Part II
Therapeutic Guidelines
Therapeutic Guidelines - IADVL
Therapy Letter
View Point
What’s new in Dermatology
View/Download PDF
Letter to Editor
doi: 10.4103/0378-6323.116744
PMID: 23974589

Role of polymerase chain reaction in detection of genital herpes

Kapil Goyal1 , Radha Kanta Ratho1 , AJ Kanwar2 , Baijayantimala Mishra1 , Mini P Singh1
1 Department of Virology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
2 Department of Dermatology, Venereology and Leprology, Postgraduate Institute of Medical Education and Research, Chandigarh, India

Correspondence Address:
Radha Kanta Ratho
Department of Virology, Postgraduate Institute of Medical Education and Research, Chandigarh - 160 012
How to cite this article:
Goyal K, Ratho RK, Kanwar A J, Mishra B, Singh MP. Role of polymerase chain reaction in detection of genital herpes. Indian J Dermatol Venereol Leprol 2013;79:705-706
Copyright: (C)2013 Indian Journal of Dermatology, Venereology, and Leprology


Genital herpes is a significant public health disease worldwide. Clinical diagnosis of genital herpes simplex virus (HSV) infection is insensitive and non-specific and requires laboratory confirmation. We evaluated the utility of polymerase chain reaction (PCR) for the diagnosis of genital herpes in 25 patients suspected of genital herpes. After taking an informed consent, swab samples were collected from the genital ulcers for Tzanck smear, antigen detection by indirect immunofluorescence (indirect IF), virus isolation in vero cell line and HSV DNA detection by targeting Glycoprotein D region by nested PCR as described previously by Read and Kurtz. [1] The blood samples were collected using aseptic conditions for antibody detection (IgG and IgM) by Enzyme-linked immunosorbent assay (ELISA). All the enrolled patients were prescribed acyclovir by the dermatologist. Positive control (tissue culture isolates of HSV-1 and HSV-2) and negative control (reaction mix containing no DNA) were included in each run to rule out false negative and false positive reactions. Among the 25 subjects, male:female ratio was 11.5:1 with a mean age of 32.8 years. Three patients were found to be human immunodeficiency virus positive on routine screening. None of the patients were reactive to Venereal Disease Research Laboratory test (VDRL). Majority (92%) of the patients presented with recurrent infection. Minute painful genital ulcers were the common presentation in each patient with multiple lesions in 80% of them. Five patients (20%) showed the presence of multinucleated giant cells in Tzanck smear examination. HSV-2 antigen was demonstrated by indirect IF in 6 (24%) patients. None of the patients showed the presence of HSV-1 antigen. The cytopathological effect (CPE) suggestive of HSV was observed on second passage in 2 (8%) samples. The CPE was confirmed by indirect IF, both the samples were positive for HSV-2. All the patients demonstrated IgG antibodies but none of the patients showed IgM specific antibodies, against HSV type 1 or 2. Following nested DNA PCR, the 272 bp amplified product was visualized in ethidium bromide stained gel showing the presence of HSV DNA in 14 (56%) samples [Figure - 1]. The sensitivity of conventional methods, i.e., Tzanck smear, type specific antigen detection and virus isolation was 35.71%, 42.85% and 14.28% respectively when compared to HSV DNA detection by PCR. The DNA detection by PCR was able to detect an additional 12 (48%) cases, which were missed by tissue culture alone and 8 (32%) cases, which were missed by indirect-IF, 9 (36%) cases, which were missed by Tzanck smear alone. Thus, overall PCR was able to detect 7 (28%) additional cases which were missed by all the conventional techniques [Table - 1].

Table 1: Test wise positivity of laboratory confirmed genital herpes type-2 patients (n=14); + (test positive), − (test negative)
Figure 1: Results of HSV PCR in representative genital herpes patients. Lane 1: Molecular marker (100 bp), Lane 2: Positive control (272 bp), Lane 3, 5, 6, 7, 8, 9, 10, 14: Positive samples, Lane 4, 11, 12, 13: Negative samples, Lane 15: Negative control

Though, virus isolation in cell culture has been considered the gold standard for diagnosis of early genital infections but has several limitations. It is less reliable in patients presenting with ulceration, crusting or reactivations. [2] In the present study prior acyclovir treatment was reported in 57.1% patients (8/14) and intermittent shedding of virus might have been missed as samples were collected at one point of time. Though all precautions were taken to maintain the cold chain during transportation of samples, but it is well known that positivity is higher during cooler weather. [3] However, nucleic acid tests are much less affected by specimen storage beyond 48 h, freezing, thawing or bacterial contamination. Presently, the patients had HSV specific IgG antibodies but none had IgM antibodies. This may be due to the effect of prior antiviral treatment before presented to sexually transmitted diseases (STD) clinic (11/25) and it is well known that specific antivirals might inhibit the production of HSV specific type 2 antibodies. [4]

Immunofluorescence is known to be a better alternate and can give results within 6-8 h but it requires expertise to visualize the slides. Published studies have shown PCR to be 4.1 times more sensitive than culture in detecting HSV infection. [3] PCR has been reported with increased sensitivity by 13.3% in specimens from vesicular lesions, by 27.4% from ulcerative lesions and by 20% from crusting lesions. [5] Though PCR has been well characterized as a method for rapid and sensitive diagnosis of HSV. However, its role has been largely confined to the investigation of suspected HSV encephalitis where it has replaced virus culture as the gold standard. The additional advantage of PCR in case of genital herpes would be the detection of the virus in subclinical episodes of virus shedding and in undiagnosed symptomatic genital lesions. The major limitation of the present study is small sample size and large prospective studies are required to replace virus culture by PCR as the gold standard in diagnosis of genital herpes.

Read SJ, Kurtz JB. Laboratory diagnosis of common viral infections of the central nervous system by using a single multiplex PCR screening assay. J Clin Microbiol 1999;37:1352-5.
[Google Scholar]
Corey L. The current trend in genital herpes. Progress in prevention. Sex Transm Dis 1994;21:S38-44.
[Google Scholar]
Wald A, Huang ML, Carrell D, Selke S, Corey L. Polymerase chain reaction for detection of herpes simplex virus (HSV) DNA on mucosal surfaces: Comparison with HSV isolation in cell culture. J Infect Dis 2003;188:1345-51.
[Google Scholar]
Bryson YJ. Current status and prospects for oral acyclovir treatment of first episode and recurrent genital herpes simplex virus. J Antimicrob Chemother 1983;12 Suppl B: 61-5.
[Google Scholar]
Scoular A. Using the evidence base on genital herpes: Optimising the use of diagnostic tests and information provision. Sex Transm Infect 2002;78:160-5.
[Google Scholar]

Fulltext Views

PDF downloads
Show Sections