Vesicular Exanthema Research Articles

The Complete Genome Sequence of the San Miguel Sea Lion Virus-8 Reveals that It Is Not a Member of the Vesicular Exanthema of Swine Virus/San Miguel Sea Lion Virus Species of the Caliciviridae.

The complete genome sequence of the San Miguel sea lion virus-8 (SMSV-8) was determined in this study. A comparison of this sequence to other calicivirus sequences in GenBank showed that this virus is genetically distinct from the vesicular exanthema of swine virus/San Miguel sea lion virus (VESV/SMSV) strains and belongs to a novel clade within the Vesivirus genus.

Circulation of Coxsackievirus A6 in hand-foot-mouth disease in Guangzhou, 2010-2012.

BackgroundHand, foot and mouth disease (HFMD) is usually caused by Enterovirus 71(EV71), and Coxsackievirus A16 (CV-A16) in Guangzhou, the biggest city of South China. However, Coxsackievirus A6 (CV-A6) were observed increased dramatically from 2010–2012.MethodsIn order to understand and to describe the epidemiologic and genetic characteristics of CV-A6, specimens of 5482 suspected HFMD cases were collected and examined by real-time fluorescence PCR. All samples positive for enteroviruses were analyzed by descriptive statistics. Phylogenetic analysis of CV-A6 based on the VP1 sequences was performed to investigate molecular and evolutionary characteristics.ResultsCoxsackievirus A6 increased dramatically from 9.04% in 2010 to 23.21% in 2012 and became one of the main causative agents of HFMD in Guangzhou. CV-A6 attack rates were highest in one to two year olds (33.14%). Typical clinic symptoms of CV-A6 HFMD include fever (589/720, 81.81%), maculopopular rash and vesicular exanthema around the perioral area (408/720, 56.66%), intraoral (545/720, 75.69%), the buttock (395/720, 54.86%), the trunk (244/720, 33.89%), the knee (188/720, 26.11%), and the dorsal aspects of hands (437/720, 60.69%). Phylogenetic analysis showed the CV-A6 isolates in this study belonged to Cluster A1 and were similar to those found in Shanghai in 2011 and 2012 (JX495148, KC414735), Shenzhen in 2011 (JX473394), Japan in 2011 (AB649243, AB649246), France in 2010(HE572928), Thailand in 2012(JX556564) and Israel in 2012 and 2013(.KF991010, KF991012).Electronic supplementary materialThe online version of this article (doi:10.1186/1743-422X-11-157) contains supplementary material, which is available to authorized users.

Dermatological Spectrum of Hand, Foot and Mouth Disease from Classical to Generalized Exanthema

Hand, foot and mouth disease (HFMD) is classically defined as a childhood fever accompanied by a rash with vesicles or erosions of the oral mucosa, hands, feet and sometimes the buttocks. Severe neurological complications are associated with enterovirus 71 outbreaks in Asia. Recently, it has been suggested that HFMD is related to coxsackie virus A6 (CV-A6) when there is an atypical rash. The objective of the study is to determine the dermatological pattern of HFMD and to identify the virus serotypes associated with a specific dermatological pattern. A prospective, cross-sectional study was conducted in 7 pediatric dermatology units in France from March 2010 to February 2012. All children with clinically suspected diagnosis of HFMD were included. Clinical data were collected and swabs from the nasopharynx and vesicles were taken for reverse transcription polymerase chain reaction and genotyping. Only children with confirmed HFMD--defined by clinical diagnosis of HFMD and positive enterovirus polymerase chain reaction results--were included for analysis. One hundred and four children consulted for suspected HFMD, including 89 (mean age: 25.7 months; sex ratio M/F 1.54) with confirmed HFMD. Seventy-eight (87.6%) had skin lesions on sites other than hand, feet and mouth. Thirty-seven (41.5%) had 5 or more anatomical sites involved (hand, feet and mouth, buttocks, legs, arms and trunk) considered as widespread exanthema. Widespread vesicular exanthema was observed with both CV-A6 and CV-A16. Peri-oral rash was associated with CV-A6 (P < 0.001). HFMD has a clinical spectrum ranging from classical to generalized vesicular exanthema. Generalized and atypical exanthema were observed with both CV-A6 and CV-A16 infections. CV-A6 is associated with peri-oral rash.

Deteksi Penyebab dan Sebaran Kasus Kejadian Luar Biasa Hand Foot and Mouth Diseases (HFMD) Tahun 2008-2012

Hand, foot, and mouth disease (HFMD) is a common disease caused by human enterovirus (HEV). The symptoms and signs of infection among children typically present with vesicular exanthema on the soles of their feet, the palms of their hands and in their mouths, causing discomfort and feeding difficulties. The disease is caused by mainly Coxsackie virus A16 (CVA16) serotype. It has recently been associated with EV-71 serotype which can also cause outbreak and mortality. Currently the data of HFMD cases in Indonesia is not available yet therefore this study aims to identify and determine the distribution of HFMD cases in Indonesia from the outbreak. HFMD can be identified by RT-PCR that recognize PAN-EV gene in the first place and has to be continued to EV-71 in VP1 area. Besides using PCR, we use isolation method using tissue cultures which are RD and 293 cell linesrom. From 48 suspected cases were received in virology lab CBBTH, NIHRD. 26 out of 48 cases (54%) were caused by enterovirus and 3 out of 26 were EV-71 (6,25%). The HFMD cases distribute in all group of ages and sex but children under five is the most susceptible. Key words: HFMD, Enterovirus, Outbreak, EV71

Cervical epidural infusion for the treatment of herpes zoster pain. Case report with review of the literature

IntroductionHerpes zoster is caused by a reactivation of the varicella zoster virus. Clinically, the disease manifests with pain in the area of a dermatome associated with vesicular exanthema. The most common complication is post-herpetic neuralgia, more frequent among elderly patients. The treatment focuses on the control of the acute infection, relief of acute pain, and prevention of chronic pain. First line treatment is pharmacological using antiviral and analgesic drugs, and epidural injections have been used for the prevention of post-herpetic neuralgia. ObjectiveThe objective of this article is to present a review of the literature on the basis of a case report of a patient diagnosed with herpes zoster who was treated with cervical epidural infusions for 16 days. Materials and methodsSearch conducted in PUBMED, including meta-analyses, systematic reviews, clinical trials, narrative reviews and case series published between 1995 and 2012, in order to perform a non-systematic narrative review in reference to a case report. ResultsOverall, 31 articles were selected for an update of the topic proposed, and a case report was described. ConclusionsThe use of epidural infusions in patients with herpes zoster is a treatment option for acute pain; it favors passive motion of the affected limb preventing atrophy, helps prevent post-herpetic neuralgia, is available in our setting, and has a low incidence of complications.

Infusión epidural cervical para tratamiento del dolor por herpes zoster. Reporte de caso con revisión temática

IntroductionHerpes zoster is caused by a reactivation of the varicella zoster virus. Clinically, the disease manifests with pain in the area of a dermatome associated with vesicular exanthema. The most common complication is post-herpetic neuralgia, more frequent among elderly patients. The treatment focuses on the control of the acute infection, relief of acute pain, and prevention of chronic pain. First line treatment is pharmacological using antiviral and analgesic drugs, and epidural injections have been used for the prevention of post-herpetic neuralgia. ObjectiveThe objective of this article is to present a review of the literature on the basis of a case report of a patient diagnosed with herpes zoster who was treated with cervical epidural infusions for 16 days. Materials and methodsSearch conducted in PUBMED, including meta-analyses, systematic reviews, clinical trials, narrative reviews and case series published between 1995 and 2012, in order to perform a non-systematic narrative review in reference to a case report. ResultsOverall, 31 articles were selected for an update of the topic proposed, and a case report was described. ConclusionsThe use of epidural infusions in patients with herpes zoster is a treatment option for acute pain; it favors passive motion of the affected limb preventing atrophy, helps prevent post-herpetic neuralgia, is available in our setting, and has a low incidence of complications.

Development and evaluation of multiplex RT-LAMP assays for rapid and sensitive detection of foot-and-mouth disease virus

This paper describes the evaluation of four novel real-time multiplex reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for rapid and sensitive diagnosis of foot-and-mouth disease (FMD). In order to overcome the genetic diversity of FMD viruses (FMDV), these multiplex RT-LAMP assay pairs were established by combining four newly designed primer sets with two primer sets that had been previously published. Using a real-time turbidimeter to detect amplification products and a panel of 300 samples collected throughout the world over a 78-year period, the performance of the multiplex RT-LAMP assays was compared with a FMDV-specific real-time RT-PCR assay. The most successful of the four multiplex RT-LAMP assays achieved a diagnostic sensitivity and specificity of 98.0% and 98.1%, and did not falsely detect FMDV in known negatives or samples containing swine vesicular disease virus, vesicular stomatitis virus or vesicular exanthema of swine virus. Furthermore, the analytical sensitivity of this multiplex RT-LAMP assay was at least as good as the individual component RT-LAMP tests. This is the first report of the development of a multiplex RT-LAMP to accommodate the high sequence variability encountered in RNA virus genomes and these results support the use of RT-LAMP as a cost-effective tool for simple diagnosis of FMD.

Hand, Foot, and Mouth Disease Outbreak and Coxsackievirus A6, Northern Spain, 2011

Hand, Foot, and Mouth Disease Outbreak and Coxsackievirus A6, Northern Spain, 2011

886 Varicella Complicated with Lobar Pneumonia and Parapneumonic Pleurisy

Background and AimsChickenpox is an acute, benign, highly contagious disease characterized by generalized vesicular exanthema with self-limited evolution. Pneumonia is the most serious complication of varicella, occurring more frequently in...

Epitope Insertion at the N-Terminal Molecular Switch of the Rabbit Hemorrhagic Disease Virus T=3 Capsid Protein Leads to Larger T=4 Capsids

Viruses need only one or a few structural capsid proteins to build an infectious particle. This is possible through the extensive use of symmetry and the conformational polymorphism of the structural proteins. Using virus-like particles (VLP) from rabbit hemorrhagic disease virus (RHDV) as a model, we addressed the basis of calicivirus capsid assembly and their application in vaccine design. The RHDV capsid is based on a T=3 lattice containing 180 identical subunits (VP1). We determined the structure of RHDV VLP to 8.0-Å resolution by three-dimensional cryoelectron microscopy; in addition, we used San Miguel sea lion virus (SMSV) and feline calicivirus (FCV) capsid subunit structures to establish the backbone structure of VP1 by homology modeling and flexible docking analysis. Based on the three-domain VP1 model, several insertion mutants were designed to validate the VP1 pseudoatomic model, and foreign epitopes were placed at the N- or C-terminal end, as well as in an exposed loop on the capsid surface. We selected a set of T and B cell epitopes of various lengths derived from viral and eukaryotic origins. Structural analysis of these chimeric capsids further validates the VP1 model to design new chimeras. Whereas most insertions are well tolerated, VP1 with an FCV capsid protein-neutralizing epitope at the N terminus assembled into mixtures of T=3 and larger T=4 capsids. The calicivirus capsid protein, and perhaps that of many other viruses, thus can encode polymorphism modulators that are not anticipated from the plane sequence, with important implications for understanding virus assembly and evolution.

Seneca Valley Virus and Vesicular Lesions in a Pig with Idiopathic Vesicular Disease

Swine Idiopathic Vesicular Disease (SIVD) syndrome is characterized by the formation of ulcers, erosions and vesicles on the skin, coronary bands and in the oral cavity of pigs. The clinical importance of SIVD is its resemblance with vesicular foreign animal diseases. Although the etiology of SIVD remains unknown, Seneca Valley virus, which belongs to the family Picornaviridae, was previously identified in such pigs. Here, we report gross and histopathologic findings in a 6-month-old intact male Chester White boar presented with a history of anorexia, lethargy and lameness.Intact and ruptured vesicles, erosions and ulcers were clinically observed within the oral cavity, around the nares, coronary bands and all four limbs. Various diagnostic tests were negative for swine vesicular disease virus, footand- mouth disease virus, vesicular exanthema of swine virus and vesicular stomatitis virus infection. However, vesicular scrapings and oral pharyngeal fluid were positive for the presence of Seneca Valley virus by RT-PCR.

Multiplex RT-PCR detection and microarray typing of vesicular disease viruses

A vesicular disease multiplex reverse transcription (RT)-PCR with an accompanying microarray assay was developed for simultaneous detection and typing of foot-and-mouth disease virus (FMDV) and vesicular stomatitis virus (VSV), and for the detection of swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV). The multiplex RT-PCR successfully detected viral RNA from a collection of 49 strains of vesicular viruses, including multiple strains from all seven serotypes of FMDV and both serotypes of VSV. The multiplex RT-PCR was also able to produce amplified products from the RNA genome of all four viruses simultaneously in mixed samples. An indirect (post-PCR labelling) amplicon labelling method and a direct (concurrent labelling with PCR) amplicon labelling method were compared for the purpose of microarray detection and typing. Accurate detection and typing was achieved with all strains tested in the microarray assay which utilized 163 virus- and serotype-specific probes. It was observed that microarray increased detection for some samples compared to using multiplex RT-PCR alone. This was most likely due to signal amplification resulting from fluorescent labelling. The limit of detection of the microarray assay was as low as 4.6 TCID 50/mL for FMDV. No amplification products or microarray reactivity was observed with non-target livestock pathogens tested or with samples collected from healthy cattle, sheep and pigs. All FMDV and VSV serotypes were detected as early as 2 days post-inoculation from oral swabs obtained from cattle infected experimentally.

Pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay

A novel assay for the pan-serotypic detection of foot-and-mouth disease virus (FMDV) was designed using a 5′ conjugated minor groove binder (MGB) probe real-time RT-PCR system. This assay targets the 3D region of the FMDV genome and is capable of detecting 20 copies of a transcribed RNA standard. The linear range of the test was eight logs from 2×101 to 2×108 copies and amplification time was approximately 2h. Using a panel of 83 RNA samples from representative FMDV isolates, the diagnostic sensitivity of this test was shown to be equivalent to a TaqMan real-time RT-PCR that targets the 5′ untranslated region of FMDV. Furthermore, the assay does not detect viruses causing similar clinical diseases in pigs such as swine vesicular disease virus and vesicular stomatitis virus, nor does it detect marine caliciviruses causing vesicular exanthema. The development of this assay provides a useful tool for the differential diagnosis of FMD, potentially for use in statutory or emergency testing programmes, or for detection of FMDV RNA in research applications.

A simple and rapid colloidal gold-based immunochromatogarpic strip test for detection of FMDV serotype A

A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test, affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip, the capture antibody was laid on a sample pad, the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7 ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C, Swine vesicular disease (SVD), Vesicular stomatitis virus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically, the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple, easy and fast for clinical testing on field sites; no special instruments and skills are required, and the result can be obtained within 15 min. To our knowledge, this is the first rapid immunochromatogarpic assay for serotype A of FMDV.

Index of Suspicion in the Nursery

A woman presents with decreased fetal movement, and ultrasonography reveals hydrops fetalis. Specific findings include massive ascites and a small pericardial effusion that was not present on ultrasonography 1 week ago. A 37 weeks' gestation boy is delivered via emergent cesarean section. Apgar scores are 7 and 9 at 1 and 5 minutes, respectively. The baby is intubated endotracheally 10 minutes after delivery because of increased work of breathing from his markedly distended abdomen and transported to the neonatal intensive care unit (NICU). Physical examination findings are normal, except for increased work of breathing and a tight, distended abdomen with visibly dilated veins. Initial laboratory results include a white blood cell count of 18.3×103/mcL (18.3×109/L), hemoglobin of 9.7 g/dL (97 g/L), and platelet count of 331×103/mcL (331×109/L). Chemistry results are within normal limits. Blood is sent for cultures, and empiric therapy with ampicillin and gentamicin is initiated. The baby also receives a 15-mL/kg packed red blood cell transfusion. A Replogle tube is placed to low intermittent suction. The infant is made NPO and started on intravenous fluids. Paracentesis is performed to relieve the pressure on his diaphragm and improve breathing, and 755 mL of green/brown fluid with particulate matter that has the appearance of meconium is aspirated. Umbilical lines are placed and pediatric surgery is consulted. Abdominal radiography shows intra-abdominal calcifications. An 11-day-old term girl whose mother had a positive cervical culture for human papillomavirus (HPV) is referred for admission because of a rectal temperature of 38.6°C and a history of “gasping for air.” Four days after birth, the baby became fussy, especially with feedings, and developed an intermittent cough and hoarse cry. Over several days, she became sleepier, only waking every 4 to 5 hours to breastfeed. On the pediatric ward, the baby …

The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus

BackgroundPrompt detection of PRRSV in the field samples is important for effective PRRS control, thereby reducing the potentially serious economic damage which can result from an outbreak. In this study, a rapid SYBR-based, one step real-time RT-PCR quantitative reverse transcription PCR (qRT-PCR) has been developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). Primers were designed based on the sequence of highly conservative region of PRRSV N gene.ResultsThe sensitivity of the real-time qRT-PCR assay was achieved through PRRSV ch-1a RNA for the generation of a standard curve. The detection limit of the assay was found to be 9.6 RNA copies per reaction mixture. This assay had excellent intra- and inter-assay reproducibility as in total 65 field samples were screened for the presence of PRRSV by conventional RT-PCR in parallel with qRT-PCR, and the detection rate increased from 60.0% to 76.9%. Moreover, the specificity result indicated that this assay could reliably differentiate PRRSV from the other swine viral diseases, such as classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV).ConclusionThe real-time qRT-PCR assay described in this report allows the rapid, specific and sensitive laboratory detection of PRRSV in field samples.

The Development of a Multiplex Reverse Transcription Polymerase Chain Reaction for Detection of Porcine Reproductive and Respiratory

A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). A set of three pairs of primer were designed based on the sequence of high conservative region of PRRSV. The diagnostic accuracy of the multiplex RT-PCR assay was evaluated using 25 field clinical samples, 10 reference strains and 6 true-negative samples. Simultaneously, the specificity and sensitivity of this method were evaluated. The result indicated that this assay could reliably differentiate between PRRSV and other swine viral disease, such as classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV). Meanwhile, this assay was shown to be 10-fold more sensitive than the conventional single RT-PCR (conventional sRT-PCR) method. The results indicated that the multiplex RT-PCR developed in this study has high sensitivity, strong specificity and easy operation,it is not only suitable for diagnosis of samples of insignifiance content,but also it has important applied value in getting rid of animals with recessive virus and veterinary quarantine .The method may provide a new avenue to the rapid detection of this important pathogen in one reaction.

Characterization of San Miguel Sea Lion Virus populations using pyrosequencing-based methods

San Miguel Sea Lion Virus (SMSV) is a small RNA virus in the genus Vesivirus with an unusually broad host range. Three populations of SMSV were examined by PCR amplification of the capsid precursor and putative helicase genes, followed by pyrosequencing. The populations were nasal swabs from two SMSV infected California sea lions (Zalophus californianus) from two different years, and a virus isolate from the earlier swab that was passaged in cell culture five times. In the capsid precursor, extensive deletions were prevalent in the passaged virus but uncommon in the clinical samples. A greater prevalence of point mutations was seen in the capsid precursor gene than in the putative helicase gene. In culture, the minority sequence in the capsid precursor at nucleotide position 5826 rapidly shifted after five passages to become the majority sequence. Levels of diversity at individual sites showed much more similarity between the two clinical samples than between the earlier clinical sample and the passaged culture from the same sample. SMSV appears to behave as a quasispecies. Assessment of original patient samples is preferable for understanding clinical SMSV populations.

Expression and self-assembly of virus-like particles from two genotypes of marine vesiviruses and development of an ELISA for the detection of antibodies

Sequences encoding the major and minor capsid proteins (VP1 and VP2) from two marine vesivirus isolates (Steller sea lion viruses V810 and V1415) were engineered for expression of virus-like particles (VLPs) in the baculovirus system. The resulting VLPs were morphologically similar to native vesivirus virions. Purified VLPs were probed in immunoblots with pooled antisera specific for nine San Miguel sea lion virus (SMSV) types, and a predominant protein of approximately 60kDa was detected. An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies was developed in which the VLPs served as antigen. The VLPs were adsorbed to the wells of a microplate, and the specificity of the ELISA was established with hyperimmune sera raised against 24 serotypes of the genus Vesivirus. The ELISA was used to screen for the presence of vesivirus specific antibodies in the sera of free-ranging Steller sea lions. The ELISA results demonstrated that Steller sea lions that inhabit the Pacific Ocean waters of southeast Alaska are widely exposed to antigenically related marine vesiviruses, while no previous exposure could be demonstrated using VLP antigens in 17 Steller sea lions from the Aleutian Islands. The broad reactivity of these VLPs and their non-infectious nature will facilitate global sero-epidemiological studies aimed at determining the incidence and prevalence of marine vesiviruses in mammals that inhabit the Pacific and Atlantic oceans as well as susceptible terrestrial animals.

Development and laboratory evaluation of a lateral flow device for the detection of swine vesicular disease virus in clinical samples

A lateral flow device (LFD) for the detection of swine vesicular disease (SVD) virus (SVDV) and differential diagnosis from foot-and-mouth disease (FMD) was developed using a monoclonal antibody (Mab C70). The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of SVDV and porcine teschovirus (enterovirus; PEV). The collection of test samples included 157 which were positive for SVDV (84 vesicular epithelial suspensions and 73 cell culture antigens) from suspected cases of vesicular disease in pigs collected from 14 countries between 1966 and 2008 and 663 samples which were either shown to be negative for SVDV and FMD virus (FMDV) or else collected from healthy pigs or demonstrated to be positive for FMDV, PEV or vesicular exanthema (VEV) and collected from 16 countries between 1965 and 2008 or else were derived from experimental animals. Three further samples containing vesicular stomatitis virus (VSV) were also tested. The diagnostic sensitivity of the LFD for SVDV was similar at 82% compared to 86% obtained by the reference method of antigen ELISA, and the diagnostic specificity was 100% compared to 99.7% for the ELISA. The device recognized virus strains of each of the known genotypes of the sole SVDV serotype. Reactions with FMDV, VEV, VSV and PEV which can produce clinically indistinguishable syndromes in pigs, did not occur. These data illustrate the potential for the LFD to be used next to the animal for providing rapid and objective support to veterinarians in their clinical judgment of vesicular disease in pigs and for the specific pen-side diagnosis of SVD and differential diagnosis from FMD.