To explore the antiviral activity of nano-realgar against herpes simplex virus Type II (HSV-2) in vitro.
 Methods: Acyclovir (ACV) as a positive control, the cytotoxicity of nano-realgar at different concentrations (including 200.00, 150.00, 100.00, 50.00, 25.00, 12.50, 6.25, 3.13, 1.54, 0.78, 0.39 and 0 mg/L) on normal Vero cells were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. HSV-2 virus titer was determined by plaque assay, and the Vero cells model of HSV-2 infection was established. Subsequently, the antiviral effects of nano-realgar at different concentrations (including 20.00, 10.00, 5.00, 2.50, 1.25, 0.63, 0.31, 0.15, 0.08, 0.04 and 0 mg/L) on infected cells model were evaluated by the observation of cytopathic effect (CPE) and MTT method under the 3 modes including pre-treatment, treatment and direct inactivation.
 Results: The 50% cytotoxic concentration (CC50) of nano-realgar on Vero cells was 37.15 mg/L. The titer of HSV-2 was 7.30 log PFUs/mL. In the 3 modes, the half-maximal effective concentration (EC50) of nano-realgar on HSV-2 infected Vero cells were 0.13, 1.80 and 0.52 mg/L, and the corresponding therapeutic index (TI) were 285.77, 20.64, 71.44, respectively. The TI value of nano-realgar on pre-treatment mode was higher than that of nano-realgar on treatment and direct inactivation modes.
 Conclusion: Nano-realgar can play a good anti-HSV-2 activity in the 3 modes (pre-treatment, treatment and direct inactivation), and the anti-HSV-2 efficacy of nano-realgar on pre-treatment mode is better than that of nano-realagr on other 2 modes.
Animals ; Antiviral Agents ; Arsenicals ; Chlorocebus aethiops ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Sulfides ; Vero Cells

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*

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 development of a multiplex polymerase chain reaction (PCR) method for rapid and accurate detection and typing of herpes simplex virus type 1 (HSV-1), and type-2 (HSV-2), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) is very important for clinical diagnosis to allow the deliver of therapy as early as possible. Large scale amplifications by multiplex PCR of viral DNA can lower the cost and time for viral diagnosis. In this study, therefore sensitive quadruplex PCR was achieved by optimizing parameters such as primers, and 1.5 mM magnesium and 200 uM dNTPs concentrations. The concentrations of HSV-1, HSV-2, CMV and EBV primers were 0.5, 0.3, 0.25 and 0.25 pmoles, respectively. Optimal annealing temperature was 54 degrees C. Employing these conditions, we could detect 10 copies of reconstructed template plasmid DNA, which were cloned to vectors containing target sequences of viral DNA. PCR products of 271 bp for HSV-1, 231 bp for HSV-2, 368 bp for CMV, and 326 bp for EBV were separated on 5.0% polyacrylamide gel electrophoresis and confirmed by direct sequencing. The present study showed that the quadruplex PCR assay described herein has potential application in clinical diagnosis, when rapid, accurate detection and typing of viruses HSV-1, HSV-2, CMV or EBV are necessary.
Cytomegalovirus/classification/*isolation & purification ; Herpesvirus 1, Human/classification/*isolation & purification ; Herpesvirus 2, Human/classification/*isolation & purification ; Herpesvirus 4, Human/classification/*isolation & purification ; Human ; *Polymerase Chain Reaction/methods ; Sensitivity and Specificity ; Support, Non-U.S. Gov't

Herpes simplex virus type 2 (HSV2) meningitis primarily develops during or following a primary genital HSV2 infection that was acquired from sexual contact or through the birth canal during delivery from mother. We describe a 15 year old virgin without history of previous herpes simplex infection who developed 2 episodes of HSV2 meningitis. Although recurrent meningitis due to HSV is primarily seen in young or sexually active adults. HSV2 meningitis should be in the differential diagnosis of recurrent meningitis in adolescent patients.
Adolescent* ; Adult ; Diagnosis, Differential ; Female ; Herpes Simplex ; Herpesvirus 2, Human ; Humans ; Meningitis* ; Mothers ; Parturition ; Simplexvirus*

We describe two patients with acute myeloradiculitis associated with herpes simplex virus type 2 (HSV-2). They were previously healthy and immunocompetent and had no history of herpes infection or rash. Myeloradiculitis manifested as an acute flaccid paralysis that primarily involved the conus medullaris and cauda equina. laccid paralysis can be caused by HSV-2 myeloradiculitis, and so early antiviral treatment should be considered.
Cauda Equina ; Exanthema ; Herpes Simplex* ; Herpesvirus 2, Human* ; Humans ; Myelitis ; Paralysis* ; Radiculopathy ; Simplexvirus* ; Spinal Cord

In the present study, we have tried to isolate and identify herpes simplex virus type 2(HSV 2) from clinical specirnens, which were inoculated into Vero cell line and grown. Eight strains of viruses were isolated from 20 suspected cases diagnosed from the pr ivate clinics in Seoul. Viruses isolated from 4 rnale and 1 female cases with active lesion were identified to the HSV 2 by indirect immunofluorescence using monoclonal antibody to HSV-2. In addition, morphology of the isolated viruses were observed under electron microscope.
Female ; Fluorescent Antibody Technique, Indirect ; Herpes Simplex* ; Herpesvirus 2, Human* ; Humans ; Seoul ; Simplexvirus* ; Vero Cells

BamHI restriction patters and genomic library of Herpes simplex virus type 2 (HSV-2) stram G were constructed, and locations of the glycoproteins gB and gD, and it genes on the fragments were detected by Southern blot analysis. HISV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in PHLA2-21 and PHLA2-22 recombinant plasmids, gB gene in PHLA2-24 plasmic, and it gene in PHLA2-11 clone by Southern blot analysis.
Blotting, Southern ; Clone Cells ; DNA ; Genomic Library ; Glycoproteins* ; Herpes Simplex* ; Herpesvirus 2, Human* ; Phosphotransferases* ; Plasmids ; Simplexvirus*

BamHI restriction patters and genomic library of Herpes simplex virus type 2 (HSV-2) stram G were constructed, and locations of the glycoproteins gB and gD, and it genes on the fragments were detected by Southern blot analysis. HISV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in PHLA2-21 and PHLA2-22 recombinant plasmids, gB gene in PHLA2-24 plasmic, and it gene in PHLA2-11 clone by Southern blot analysis.
Blotting, Southern ; Clone Cells ; DNA ; Genomic Library ; Glycoproteins* ; Herpes Simplex* ; Herpesvirus 2, Human* ; Phosphotransferases* ; Plasmids ; Simplexvirus*

Humans ; Lymphohistiocytosis, Hemophagocytic ; Herpesvirus 4, Human ; Interleukin-2 ; Epstein-Barr Virus Infections/complications* ; T-Lymphocytes