BACKGROUND: Herpes simplex virus type 1 (HSV-1) is linked specifically to oral and lip infections while HSV-2 is involved in genital infections. We evaluated a recently reported nested-multiplex ploymerase chain reaction (NM-PCR) for the detection and typing of HSV and compared the results with indirect immunofluorescence (IF) after cell culture. METHODS: One hundred thirty three specimens were received from patients suspected of having clinical HSV infections. HSV was cultured by the shell vial method and stained with type specific monoclonal antibodies. NM-PCR was performed using crude samples. RESULTS: HSV was detected in 45 (33.8%) and 46 (34.6%) of the 133 specimens by IF and NMPCR, respectively. All of the HSV IF positive specimens were also positive by NM-PCR. Typing by the two methods concurred in all but two of the 45 specimens; the two specimens were typed as HSV-1 and HSN-2, respectivey, by IF, and both as HSV-1 and HSV-2 by NM-PCR. CONCLUSIONS: Our results suggest that NM-PCR is a rapid and sensitive method for the detection and typing of HSV.
Antibodies, Monoclonal ; Cell Culture Techniques ; Fluorescent Antibody Technique, Indirect* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Lip ; Simplexvirus*

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with HSV-1 and HSV-2 standard stain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of HSV-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and HSV-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, B and BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum from respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 42.6%) HSV was detected singly or mixed infection and 19 of the cases were HSV-2 and 1 case was HSV-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, B and BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.
Adult ; Arthritis ; Child ; Coinfection ; Diagnosis ; Digestion ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Herpesvirus 2, Human ; Herpesvirus 3, Human* ; Humans ; Joints* ; Mental Competency ; Polymerase Chain Reaction* ; Simplexvirus*

One-hundred-eleven herpes simplex viruses(HSV) were isolated from l09 patients who had visited the Department of Dermatology, Ophthalmology, and Gynecology of Hanyang University Hospital from 1986 through 1988, for suspected HSV lesions. The cultured viruses were classified into HSV-1 and HSV-2 by using direct and indirect immunofluorescent staining with monoclonal antibodies against HSV. In this study, HSV type 1 were predominantly involved in the HSV lesions above the waist of the patients(83.6%), and the infections caused by HSV type 2(I3.1 %) and bath HSV types(3.3%) were also noted. In contrast, the main organism of the HSV lesions below the waist were HSV-2(80.0%), and HSV-1(16.0% ) and both types(4.0%) were also involved in. The result suggested that the number of patients with HSV 1 genital herpes and/or with non-genital herpes by HSV-2 were apparently increasing. In conclusion, it is conceivable that the classification of HSV isolates might be useful for determining prognosis as well as performing epidemiologic studies of HSV infections.
Antibodies, Monoclonal ; Baths ; Classification ; Dermatology ; Epidemiologic Studies ; Gynecology ; Herpes Genitalis ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Ophthalmology ; Prognosis ; Simplexvirus*

BACKGROUND: Genital herpes is a sexually transmitted disease, which affects millions of people worldwide, and is caused by the herpes simplex virus (HSV). Recent data has shown that in a large proportion of genital herpes, there has been a shift from HSV type 2 (HSV-2) to to HSV type 1 (HS V-1) being the main cause. OBJECTIVE: Our objective was to study the types of virus and clinical patterns of patients with genital herpes in Korea METHODS: We investigated the clinical patterns and virus types of 13 patients with genital herpes using indirect immunofluorescence (IIF) after viral culture, and/or nested-multiplex polymerase chain reaction (NM-PCR). RESULTS: Of the 13 patients, HSV-1 was isolated in 2 patients (15.4%), HSV-2 in 7 patients (53.8%), and mixed infection with both HSV-1 and HSV-2 in 4 patients (30.8%). Recurrence of lesions was found to occur when the patient had HSV-2 or a mixed infection, but not the HSV-1 infection. Of 5 patients who underwent IIF and NM-PCR simultaneously, the virus was detected by NM-PCR only, not by IIF after viral culture, in 2 of the patients. CONCLUSION: HSV-1 infection as a cause of genital herpes is increasing, but recurrence is more common in HSV-2 infection. This study demonstrates that HSV-1 and HSV-2 can be detected simutaneously in the same anatomic region of genital herpes, and that NM-PCR is a more sensitive method for the detection and typing of HSV than IIF after viral culture.
Coinfection ; Fluorescent Antibody Technique, Indirect ; Herpes Genitalis* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Korea ; Polymerase Chain Reaction ; Recurrence ; Sexually Transmitted Diseases ; Simplexvirus

Herpes simplex virus (Herpesvirus hominis), a member of the herpeptoviridae family, is among the most common infectious viral pathogens in humans. Types of HSV can be subdivided into HSV-1 and HSV-2 on the basis of physiological, serologic and epidemiologic differences, although both are closely related immunologically and DNA sequence homology. HSV-1 is frequently associated with oropharyngeal, eye and skin infections, while HSV-2 is most commonly associated with genital tract infections. Characteristically after the primary infection, HSV can be linked to latency in neural tissue. Reactivation of HSV can be triggered by sunburn, fever, stress, menstruation and local trauma. Here we report a case of reactivation of herpes simplex virus type 2 in foot dorsum followed by repetitive friction due to Sandals' trap.
Base Sequence ; Female ; Fever ; Foot* ; Friction* ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Menstruation ; Reproductive Tract Infections ; Simplexvirus* ; Skin ; Sunburn

BACKGROUND: Human herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) are responsible for a plethora of human diseases, of which cutaneous and mucocutaneous infections are the most prevalent. In its most severe form, HSV infection can cause meningitis/encephalitis. We compared the Luminex ARIES HSV 1&2 assay (Luminex Corp., Austin, TX, USA), an automated sample-to-result molecular solution, to two non-automated HSV DNA assays. METHODS: A total of 116 artificial controls were used to determine the analytical performance of the ARIES assay. Controls were prepared by spiking universal transport medium (UTM) and cerebrospinal fluid (CSF) samples from patients who tested negative for HSV by an in-house HSV-1 and -2 DNA assay with reference materials (SeraCare Life Sciences, MA, USA; ZeptoMetrix Corp., MA, USA). Another 117 clinical samples were then used to compare the clinical performance of the ARIES assay with those of an in-house assay and the FTD Neuro 9 assay (Fast Track Diagnostics, Junglinster, Luxembourg). RESULTS: The analytical sensitivity (95% limit of detection) of the ARIES assay was 318 copies/mL (UTM samples) and 935 copies/mL (CSF samples) for HSV-1 strain 96 and 253 copies/mL (UTM samples) and 821 copies/mL (CSF samples) for HSV-2 strain 09. No cross-reactivity was observed in samples spiked with 14 non-HSV microorganisms. Compared with the reference result (agreement between the in-house and FTD Neuro 9 results), the ARIES assay had overall concordance rates of 98.2% (111/113) and 100% (113/113) for HSV-1 and HSV-2, respectively. CONCLUSIONS: The ARIES assay appears to be an excellent alternative for rapid detection and differentiation of HSV in skin and genital infections, meningitis, and encephalitis.
Biological Science Disciplines ; Cerebrospinal Fluid ; DNA ; Encephalitis ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Meningitis ; Real-Time Polymerase Chain Reaction* ; Simplexvirus* ; Skin

BACKGROUND: Herpes simplex virus(HSV) infections are very common viral illnesses in dermatology and they have shown a tendency to increase in prevalence and incidence, and they would seem to have become more prevalent recently. This has resulted in an increased need for the rapid diagnosis of these infections. It has also become important to recognize the types of HSV in the clinical setting, because the two types differ in their natural histories and prognoses. OBJECTIVE: The purpose of this study is to detect and type HSV DNA by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). We investigated the relationship between the clinical manifestations and type using PCR. METHODS: The study population consisted of 40 cases of HSV infections and 10 cases of varicella-zoster virus infections as negative controls. We used a pair of primers designated by Sakaoka et. al and performed restriction analysis after PCR. RESULTS: No specific amplification was observed using the varicella-zoster virus. A total of 40 patients were examined by PCR and 28 were positive. Of 17 patients with lesions which developed above the umbilicus, l0 were positive. Out of 8 patients with lesions below the umbilicus, 5 were positive. Virus genomes were detected in the 13 out of 15 patients with eczema herpeticum. CONCLUSION: The method used in this study is useful in differentiating HSV-1 from HSV-2 in fections. These data suggest that the PCR results were nearly consistent with the clinical manifestations.
Dermatology ; Diagnosis ; DNA ; Genome ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Herpesvirus 3, Human ; Humans ; Incidence ; Kaposi Varicelliform Eruption ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length ; Prevalence ; Prognosis ; Simplexvirus* ; Umbilicus

BACKGROUND: Herpes simplex virus(HSV) infections are very common viral illnesses in dermatology and they have shown a tendency to increase in prevalence and incidence, and they would seem to have become more prevalent recently. This has resulted in an increased need for the rapid diagnosis of these infections. It has also become important to recognize the types of HSV in the clinical setting, because the two types differ in their natural histories and prognoses. OBJECTIVE: The purpose of this study is to detect and type HSV DNA by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). We investigated the relationship between the clinical manifestations and type using PCR. METHODS: The study population consisted of 40 cases of HSV infections and 10 cases of varicella-zoster virus infections as negative controls. We used a pair of primers designated by Sakaoka et. al and performed restriction analysis after PCR. RESULTS: No specific amplification was observed using the varicella-zoster virus. A total of 40 patients were examined by PCR and 28 were positive. Of 17 patients with lesions which developed above the umbilicus, l0 were positive. Out of 8 patients with lesions below the umbilicus, 5 were positive. Virus genomes were detected in the 13 out of 15 patients with eczema herpeticum. CONCLUSION: The method used in this study is useful in differentiating HSV-1 from HSV-2 in fections. These data suggest that the PCR results were nearly consistent with the clinical manifestations.
Dermatology ; Diagnosis ; DNA ; Genome ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Herpesvirus 3, Human ; Humans ; Incidence ; Kaposi Varicelliform Eruption ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length ; Prevalence ; Prognosis ; Simplexvirus* ; Umbilicus

BACKGROUND: Polymerase chain reaction(PCR) was developed to diagnose viral infections such as herpes simplex virus(HSV) more rapidly and accurately than the culture technique. Nested PCR, with knowingly higher sensitivity and specificity than the conventional PCR, has been recently applied to determine HSV isotyping. OBJECTIVE: We conducted this study to compare the prevalence of each HSV isotype among patients with HSV infections by nested PCR in relation to different age groups and body sites. METHODS: Specimens were collected with cotton-swabs from the lesions, and then snap frozen, and stored at -70 degrees C in a deep freezer until processed. They were innoculated into the monolayer of Vero(African green monkey kidney) cells, and later examined for cytopathic effects. DNAs were extracted from each specimen and were amplified by using nested PCR with the primers against the genes encoding gpD of HSV-1 and HSV-2, respectively. RESULTS: Of 127 patients, who had been diagnosed as HSV infections by the culture technique, all cases were positive in the nested PCR method. The positive cases of each isotype for HSV-1, HSV-2 and mixed infections (HSV-1 & HSV-2) were 77 (61%), 36 (28%) and 14 (11%), respectively. The prevalence of infections with HSV-1 in the age group under 19 years old was significantly higher than that in the age group over 20 years old (p<0.05). The prevalence of infections with HSV-2 in the age group over 20 years old was significantly higher than that in the age group under 19 years old (p<0.05). There was a close correlation between the HSV isotypes and their predilection sites of the body, showing HSV-1 for area above the waist including lips, oral cavity, eyes, face, fingers and chest, otherwise HSV-2 and mixed infections for area below the waist including genital organs and buttocks (p<0.001). CONCLUSION: It was suggested that the prevalence of each HSV isotype in certain age groups of the patients might be somewhat different from those reported in other studies. The affinity of HSV isotypes to the specific sites of the body was reconfirmed. This study indicated that nested PCR might be a relatively rapid and cost-efficient test with very high sensitivity and specificity for HSV isotyping.
Buttocks ; Cercopithecus aethiops ; Coinfection ; Culture Techniques ; DNA ; Fingers ; Genitalia ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Lip ; Mouth ; Polymerase Chain Reaction* ; Prevalence ; Sensitivity and Specificity ; Simplexvirus* ; Thorax ; Young Adult

BACKGROUND: Herpes simplex virus (HSV) is associated with a insignificant skin lesion, keratitis, encephalitis, congenital infection, sexually transmitted disease, or cervix cancer. There are two types of serogroup, HSV-1 and HSV-2. HSV-1 makes the lesion mainly on the above-waist area and HSV-2 makes the lesion mainly on the below-waist area. To diagnose the HSV infection, immunological or cultural methods usually have been used until now. But they are not satisfactory in terms of sensitivity, specificity, and ease of application. Recently the polymerase chain reaction (PCR) was developed. Because of the exponential nature of the amplification, this method can detect extremely small amount of DNA. We compared nested PCR with cultural method for HSV detection. METHODS: We obtained 61 specimens from the lesions of oral mucosa, face, and genital area. Samples were inoculated into the monolayer from the African green monkey kidney cell(Vero). When the slide showed cytopathic effect(CPE), HSV infection was confirmed, After extracting DNA from 61 samples, we amplified HSV DNA using nested PCR with the primers against the gene encoding glycoprotein (gD) of HSV-1 and HSV-2. RESULTS: We found 632 bp band after the 1st PCR round and 271 bp band after the 2nd PCR round with HSV-1 specific primers. HSV-2 revealed 428 bp band after the 1st PCR round and 231 bp band after the 2nd PCR round. Nested PCR showed analytical sensitivity at 10(-9) g of DNA in HSV-1 and 10(-10) g of DNA in HSV-2. Viral culture was positive in 36%, nested PCR detected HSV DNA sequence in 54% of samples. Nested PCR typed HSV, HSV-1 in 67%, HSV-2 in 39%, and mixed type in 6% of PCR-positive samples. All isolates from above-waist area were HSV-1. Seventy seven percent of 13 isolates from below-waist area were HSV-2 and 38% were HSV-1. CONCLUSIONS: Nested PCR offers a rapid, simple, and sensitive test for HSV infections of skin and mucosa.
Base Sequence ; Cercopithecus aethiops ; DNA ; Encephalitis ; Glycoproteins ; Herpes Simplex* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Keratitis ; Kidney ; Mouth Mucosa ; Mucous Membrane ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Sexually Transmitted Diseases ; Simplexvirus* ; Skin ; Uterine Cervical Neoplasms