Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Dr Mohan Z Mani

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On Sep 2018




Prof. Somashekhar Nimbalkar

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Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
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Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

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Professor and Head
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Saraswati Dental College
Lucknow
On Sep 2018




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Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




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Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2011 | Month : November | Volume : 5 | Issue : 6 | Page : 1190 - 1194 Full Version

Correlation of Serology, Tissue Culture and PCR in Identification of Herpes Simplex Type-2 infection among HIV Patients


Published: November 1, 2011 | DOI: https://doi.org/10.7860/JCDR/2011/.1616
Shameem Banu A.S, Lakshmi S., Kaveri K., Jayakumar S.

MBBS, MD (Microbiology), Department of Microbiology, Saveetha Medical College & Hospital, Saveetha University, Thandalam, Kancheepuram District – 602 105 Tamil Nadu, India. BBS, MD (Microbiology), Department of Microbiology, Saveetha Medical College & Hospital, Saveetha University, Thandalam, Kancheepuram District – 602 105 Tamil Nadu, India. MBBS, MD (Microbiology), Department of Microbiology, Saveetha Medical College & Hospital, Saveetha University, Thandalam, Kancheepuram District – 602 105 Tamil Nadu, India. MBBS, MD (Microbiology), Department of Microbiology, Saveetha Medical College & Hospital, Saveetha University, Thandalam, Kancheepuram District – 602 105 Tamil Nadu, India.

Correspondence Address :
Shameem Banu A .S.
Professor & Head, Department of Microbiology,
Saveetha Medical College & Hospital, Saveetha University,
Thandalam, Kancheepuram District – 602 105
Tamil Nadu.
Phone: 9940127670
E-mail: shameembanu10@rediffmail.com

Abstract

Background: Herpes infection in Acquired Immunodeficiency Syndrome (AIDS) patients is more severe and in atypical location. Diagnosis becomes difficult as the clinical presentation tend to be atypical. There are numerous laboratory methods to identify Herpes Simplex Virus (HSV) infection and it also becomes essential as a early initiation of antiherpetic treatment has shown to decrease Human Immunodeficiency Virus (HIV) replication and acyclovir resistance. In this study we have correlated serology of HSV type 2 with tissue culture and Polymerase Chain reaction (PCR).

Materials and Methods: This study was performed in the Institute of Microbiology, Madras Medical College and the Department of Virology, King Institute of Preventive Medicine. The sample size was 60 cases all with genital lesions such as ulcers and vesicles were selected. Out of which 30 were known HIV positive and 30 were HIV negative attending out patient Sexually Transmitted Diseases Department, Government General Hospital (GH) Chennai. Thirty cases of blood donors were taken as control population from the Blood Bank, GH Chennai. Laboratory diagnostic procedures such as serological tests, virus culture, immunoflourescence and PCR were done keeping PCR as a gold standard test for diagnosing HSV infection.

Results: Out of 60 patients, 30 were HIV positive and remaining negative. Among HIV positive patients 56.6% had IgM antibody and 83.3% had IgG antibody, were as among HIV negative patients IgM were 36.6% and 70% had IgG. Control group serological parameters for IgM, IgG HSV-2 were 6.6% and 56.6% respectively. Virus isolation was positive for three samples and PCR for six samples. On comparing with gold standard test, sensitivity and specificity for serology is 100% and 59%, for tissue culture the sensitivity is 50% and specificity is 100%.

Conclusion: HSV 2 antibody detection by type specific serology kit can be used as an effective tool in screening infection both in typical and atypical presentations before initiating treatment, as antibody detection is much easier and feasible screening test when compared to PCR and tissue culture.

Keywords

Human Immunodeficiency Virus (HIV), Herpes Simplex Virus (HSV), Tissue Culture, Polymerized Chain Reaction

Introduction
Since AIDS was first recognised 25 yrs ago, remarkable progress has been made in improving the quality and the duration of life for HIV infected persons. Improved recognition of opportunistic infection disease processes, improved therapy for acute and chronic complications and introduction of chemoprophylaxis against key opportunistic pathogens. The sexual transmission of the Human Immunodeficiency Virus (HIV) is facilitated by the presence of genital ulcer disease. Herpes Simplex Virus type 2 (HSV-2) is the most common cause of genital ulcer disease in both the developed and the developing countries. HSV-2 and its potential interaction with HIV have emerged as a major public health problem for countries which are facing the global HIV -1 pandemic (1).

The transmission of HIV-1 to sexual partners may also be aided by the presence of genital ulcers. High levels of HIV-1 have been documented in the HSV lesions. Furthermore, HSV reactivates more frequently in persons with the HIV-1 infection. These findingsindicate a higher risk of HIV-1 exposure for individuals whose sexual partners have both the HIV-1 and the genital herpes infections (2).

Up to 90% of the people with the HIV infection also have the HSV-2 infection. Most of the people who are infected with HSV-2 do not know that they have the virus because its symptoms can be mild or they may be absent. The HSV-2 infection can cause recurrent sores and breaks in the skin of the genital region, which can be mild and can often go unnoticed. The HSV-2 infection also attracts immune cells which are called the CD4 T-cells to the genital region, which HIV uses to establish or pass the infection. Multiple studies have shown that frequent genital herpes recurrences increase the amount of HIV in the blood and the genital tract. The HIV virus is also shed from genital herpes ulcers and persons with such ulcers can transmit HIV to others more efficiently (3).

The herpes infections are more severe and atypical in AIDS patients. Multiple necrotic ulcers may be present in atypical locations. Theoutbreaks may progress to more extensive and persistent locations. There may be more frequent recurrences or chronic and nearly continuous ulcerations as the HIV related immunosuppresion progresses. Patients with the human immunodeficiency virus (HIV) infection who experience the first or recurrent HSV-2 episodes can develop severe and extensive lesions, which may become difficult to control by the standard antiviral therapy (4). In some cases, a visceral spread can also occur (2).

The detection and the treatment of sexually transmitted diseases such as genital herpes actually decrease the rate of the HIV infection. The treatment of the HSV infection in HIV seropositive patients suppresses the bursts of the HIV replication which occur during the active herpes infection. All the five trials demonstrated significant reductions in both the genital and plasma HIV replication after one to three months of daily treatment with either valacyclovir or acyclovir (5). As the presentation tends to be atypical, a diagnosis which is based on clinical grounds becomes difficult and so laboratory diagnostic methods become essential, as initiating the antiherpetic treatment has been shown to decrease both the HIV replication and the emergence of acyclovir resistance in these patients.

Various tests are available for diagnosing HSV-2 infections in the laboratory, with many pros and cons. In this present study, we correlated PCR, tissue culture and serology tests to find out an ideal and feasible tool for diagnosing the HSV type 2 infection.

Material and Methods

Study Participants and Design
This study was performed at the HIV Reference Centre, Institute of Microbiology, Madras Medical College, Chennai and in the Department of Virology, King Institute of Preventive Medicine, Guindy. The study group was the patients who attended the Out Patients Sexually Transmitted Diseases (STD) Department, Government General Hospital, Chennai and the control population was obtained from the Blood Bank, Government General Hospital, Chennai.

The study population represented both the male and female patients who attended the STD clinic. The patients were interviewed regarding their demographic characteristics, their medical history, their HIV/AIDS status knowledge, and their clinical symptoms. In brief, all the patients received pre-and post-test counseling for HIV and those who gave consent for the study were included.All the participants were informed about the confidentiality of their test results. PCR was the gold standard test, with which we compared tissue culture and serology.

Specimen Collection and Laboratory Tests
The sample size was 60 cases (with vesicles or ulcer) and 30 controls. Blood specimens which were obtained by venipuncture were tested for HIV, IgM and IgG for HSV-2. Sterile cotton swabs were used to collect scrapings from the base of the vesicular and the ulcerative lesions for the virus isolation and the PCR assays.

Serologic testing for HIV:
The serum samples were screened by using a commercially available EIA kit for the identification of the HIV-1 and the HIV-2 antibodies (LAB Systems HIV-EIA, an indirect, solid phase immunoassay).

Serologic testing for HSV-2:
(6) The serologic testing for HSV-2 was performed on all the specimens by using IgM type specific ELISA (NovumHSV2 IgM immunoassay) and Herpes simplex virus type 2 IgG ELISA (NovumHSV2 IgG immunoassay). Positive andnegative control serum samples were used in each experiment. The cut-off was determined by dividing the optical density (OD) of the positive and the negative controls. The average absorbance (OD) value of the cut-off serum which was run in duplicate was calculated. The cut off index (COI) of each serum sample was determined by dividing the OD which was obtained for that serum sample, by the average OD of the cut-off serum. A COI below or equal to 1 was considered to be negative, that above 1.1 was considered to be positive and a COI between 1 and 1.1 was considered as borderline.

Preparation of the Specimen for the Virus Isolation and for PCR Amplification
A swab was immediately transferred into 2ml of viral transport medium and transported to the laboratory in a cold chain. The specimen was processed by using a standard protocol and it was inoculated immediately for culture. The remaining was stored at -70 deg C for PCR testing later.

Virus Isolation (7) 0.2 ml of each of the processed samples was inoculated into tubes which contained a confluent verocell monolayer (Vervet monkey kidney cell lines in Minimal Essential Medium which was supplemented with 2% foetal calf serum) for virus isolation by using standard tissue culture techniques. The cultures were investigated daily by using an inverted microscope to see the cytopathological effect (CPE) for a week.The cytopathic effects which were produced by the Herpes Simplex virus were ballooning of the infected cells and the formation of multinucleated giant cells. The tubes which showed the cytopathic effect were confirmed by immunoflourescence. Staining was done with fluorescent iso thiocanate which was conjugated to HSV2 polyclonal antiserum (reagent obtained from J. Mitra). Prototype Herpes Simplex 2 strains were used as positive controls and uninfected cell cultures were used as negative controls. Specific fluorescence was detected.

PCR Amplification
(8) The stored, processed samples were centrifuged and 50μl of the supernatant was added to an equal volume of DNA extraction buffer. The DNA extraction was done by suing a NeoDin HSV Screen and Type I, II PCR Kit-One tube nested PCR kit. The target region was the nucleotides and the nucleosides of the gD gene of Herpes Simplex from the clinical samples. The PCR amplified product was electrophoresed and the product size was analyzed by using a UV transilluminator. The product size of the sample was compared with the positive and negative controls which were provided in the kit and also with the molecular weight markers.

Results

Ninety subjects were enrolled in the study, of which 30 were HIV positive (20 males and 10 females) and 30 were HIV negative individuals (19 males and 11 females) who attended the STD clinic and 30 cases (14 males and 16 females) attended the Blood Bank, which were taken as the control group.

In the HIV positive study group, the incidence of the genital lesions was 50% in the 20-29 years age group and it was 50% in the 30- 39 years age group. Out of the 60 patients, 50 patients presented with ulcers (22 - HIV positive and 28 - negative) and 10 patients with vesicles (8- HIV positive and 2- HIV negative).

The seroprevalence of the HSV-2 antibody (IgM, IgG) among the HIV positive and negative patients as compared to the control group is shown in (Table/Fig 1).

The virus was isolated only in 3 cases (5%), for which the cytopathic effect was confirmed by direct immunoflourescence.

A total of 6 cases (10%) were positive for PCR. The positivity results for the virus isolation and for PCR among the HIV cases with vesicles and ulcers with respect to the treatment is shown in (Table/Fig 2).

The sensitivity and the specificity for serology and tissue culture were computed by using the 2 by 2 table as shown in (Table/Fig 3). The statistical analysis for the sensitivity and specificity of these two tests was done by using the Receiver Operating Characteristic (ROC) curve. The value for the serology of IgM by using the ROC curve was 0.796 and for tissue culture, it was 0.750. The ROC curve is shown in (Table/Fig 4).

Discussion

The HIV-1 epidemic in India is now 24 years old and it has spread rapidly across the country; as a result, India has the potential to have more HIV-1–infected individuals than any country in the world (1). Among all the causes, HSV-2 is the predominant cause of genital herpes, accounting for approximately 85% of the cases of primary genital herpes and >95% of the cases of recurrent genital herpes in Sydney, Australia, and elsewhere (9).

The importance of the association between genital herpes and the HIV-l infection is underscored by the fact that the prevalence of genital herpes is much greater than the prevalence of syphilis or chancroid. Therefore, the risk of acquiring HIV-1 was increased to 2-fold in the patients with genital herpes (10).

In addition, most of the patients with genital herpes are likely to have recurrent episodes of genital ulceration. Thus, if genital herpes infections are the risk factors for the acquisition of HIV-1, they have the potential to contribute to the continued spread of the HIV-1 infection because of their recurrent nature, due to the high prevalence of genital herpes in populations who are at a risk for acquiring the HIV-1 infection and due to the large numbers of herpes-infected individuals who continue to engage in sexual activities despite their infections.

There are several possible biological mechanisms that HSV-2 could use to act as a cofactor in HIV acquisition or transmission. First, the HSV-2 reactivations result in mucosal or epithelial disruption, creating a portal of exit or entry for HIV, to which the activated CD4 cells are recruited. There also appear to be several cellular interactions that promote the establishment of the HIV infection; coinfection with HSV-2 may lead to the creation of “pseudotypes” (i.e., particles containing the HIV viral genome which are enveloped in the HSV surface glycoprotein), thus allowing HSV to infect the cells that could not be infected by HIV alone (11). The HSV-2 infection may also promote the increased expression of the HIV target cells (i.e., the CCR5+ CD4 cells and the immature dendritic cells) (12).

The episode of acute HSV infection increases HIV transcription, as was evidenced by the intracellular HIV gag messenger RNA. Co-infection of the human CD4 cells with HSV and HIVresults in a undirectional, accelerated replication of HIV. The prevalence of HSV-2 shedding is greater in HIV seropositive individuals and it increases significantly with a decrease in the CD4 count.

There are also several mechanisms that may explain how HSV-2 increases the levels of genital and plasma HIV, thus enhancing the HIV transmission. In coinfected individuals, the HSV-2 proteins may increase the replication of HIV at the mucosal sites by transactivation of the HIV long terminal repeat (13). Cytokine release may also stimulate HIV replication. A recent study demonstrated that coinfection with HIV could lead to a depletion of the immune cells which are responsible for controlling the HSV-2 reactivation, resulting in an impaired immune control of HSV-2, and leading to further HSV-2 reactivation and subsequent increases in the HIV levels in the genital tract (12).

Some data suggest that the lesions which are caused by HSV may be atypical in morphology as compared to those which have been clinically described, thereby delaying or preventing an accurate diagnosis (8). The atypical manifestations include vulvar, penile, or perianal fissures, localized erythema, and back pain without genital lesions. Both of these groups are at risk of transmitting the virus. The HSV-2 virus is capable of causing either genital ororopharyngeal infection and can produce lesions that are clinically indistinguishable from lesions which are produced by the HSV 1 virus. Extra genital lesions are more common in women than in men. The areas which are generally involved are the buttock, groin, thigh and the fingers. The causes of the extra genital lesions are the autoinoculation of the virus, viral reactivation in another part of the dermatome and viraemic spread (2).

In this present study, the seropositivity for HSV 2 was more in the HIV positive group as compared to that in the HIV negative group (56.6% and 36.6%). This was similar to the results which were obtained in a study which was conducted at the John Hopkins University School of Medicine, Baltimore (73% and 48.9) (14).

The 28 patients who had presented primarily with vesicular and ulcerative lesions had IgM antibodies and 14 of them had IgG antibodies also. But the patients with recurrent vesicles and ulcerative lesions had IgG antibodies without the IgM antibodies.

In the control population, the IgM positivity percentage was 6.6%. The seropositivity in the control group was well supported by numerous studies which had revealed that a majority of the HSV-2 infections were left undiagnosed. In the large NHANES survey, only 9% of the persons with the HSV-2 antibodies had knowledge about their disease. Conversely, 22% of those who denied a history of genital herpes were found to be HSV-2 seropositive (2).

It has been estimated that about 20% of the patients with HSV-2 antibodies are truly asymptomatic or that they have lesions only in locations such as the cervix, that are impossible to observe (15). The remaining 60% of the undiagnosed persons with genital HSV-2 infection have symptoms that are not recognized by either the physicians or the patients as being caused by herpes (2). The sensitivity of antibody detection of our ELISA kit was 100%, which well correlated with the findings of a study which was done in Africa (16), where they compared various ELISA kits and found that their sensitivities ranged between 83.3 and 100%. The differences in the specificity were more marked, with estimates of between 46.6 and 97.7%. The detection of antibodies allows the diagnosis of an infection when virological methods such as culture, antigen detection, or PCR are impractical (17).

The observation which was made from this study was that, PCR (10%) was able to pick up more cases than virus isolation (5%), which was supported by Safrin et al (8) studies wherein the positivity for PCR was (50%) and that for virus isolation was (39.8%).

Moreover, in the present study, the virus was isolated (5%) from the samples which were taken from the patients with vesicular lesions, who had not initialized the antiherpetic treatment. Virus isolation was not present in the remaining cases and this was probably due to the decrease in the viral shedding after the treatment. There was no virus isolation from the ulcerative lesions with and without treatment, and this can be explained by the fact that the virus shedding from the ulcers was not consistent, whereas PCR was able to pick up cases from both the vesicles and the ulcers (10%) and even from cases where the antiherpetic treatment had been initiated. Our finding of the greater sensitivity of PCR than virus isolation for the older or crusted lesions, mirrors that of Cone et al, who showed that the mean duration of the HSV DNA positivity was more than two fold longer than of that of the virus culture (18). Although viral culture has been widely used as a means to confirm the clinical diagnosis of the HSV infections, it has been shown to lack sensitivity when the lesion is partially healed or crusted. Improper handling of the specimen could further decrease the yieldof the virus culture (17).

The selection of an effective diagnostic technique depends on a number of parameters like the clinical stages of presentation and the number of days after the clinical illness, whether it was a primary or recurrent case and whether any antiherpetic treatment had been taken or not. If a case presented within the initial 14 days with vesicles and without taking any antiviral therapy, then virus isolation could be done. On the contrary, if a case presented with ulcers and if antivirals had already been initiated for it, then PCR would be ideal.

HSV 2 antibody detection by using a type specific serology kit can be used as an effective tool in screening the infection, both in typical and atypical presentations before initiating the treatment, because it is a much easier and feasible screening test as compared to PCR and tissue culture. The most significant potential application of serology is to detect the silent carriers of HSV-2, especially in high-risk settings such as STD clinics (2). The PCR and tissue culture techniques need a good infrastructure and the availability of trained personnel.

In cases with a high clinical suspicion, with the atypical presentation of the HSV-2 infection, IgM antibody detection can be done to initiate the empirical antiviral treatment which can help in preventing the spread of the virus. Moreover, various diagnostic methods alone cannot halt the spread of genital herpes; it is an important step in the clinical management and counseling of the patients who wish to have full information about the status of their sexual health.

Key Message

Clinical presentations of Herpes Simplex Virus (HSV) infection tend to be atypical in HIV patients. Laboratory screening test or diagnostic test for Herpes Simplex Virus (HSV) infection is essential before initiating antiherpetic treatment.

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