Suppression of CD80 Expression by ICP22 Affects Herpes Simplex Virus Type 1 Replication and CD8+IFN-γ+ Infiltrates in the Eyes of Infected Mice but Not Latency Reactivation.

Previously, we reported that herpes simplex virus type 1 (HSV-1) ICP22 binds to the CD80 promoter and suppresses its expression in vitro and in vivo. To better understand the impact of ICP22 binding to CD80 on HSV-1 infectivity and pathogenicity, we mapped the region of ICP22 required to bind the CD80 promoter to a 40-amino-acid (aa) region of ICP22. We constructed a recombinant HSV-1 expressing a truncated form of ICP22 that lacks these 40 aa, which does not bind to the CD80 promoter (KOS-ICP22Δ40) and retains the ability to replicate efficiently in rabbit skin cells, in contrast to ICP22-null virus. The replication of this recombinant virus in vitro and in vivo was higher than that of the ICP22-null virus, but virus replication kinetics were lower than those of the wild-type (WT) control virus. Similar to ICP22-null virus, the KOS-ICP22Δ40 mutant virus increased CD80 expression in dendritic cells (DCs) and interferon gamma (IFN-γ) expression in CD8+ T cells but not CD4+ T cells in infected mouse corneas. In contrast to the significantly reduced virus replication in the eyes of ocularly infected mice, the levels of latency reactivation were similar between KOS-ICP22Δ40 virus and WT virus. Thus, blocking ICP22 binding to the CD80 promoter using a recombinant virus expressing a truncated ICP22 that lacks CD80 promoter binding appears to reduce virus replication and enhance CD8+IFN-γ+ infiltrates in corneas of infected mice, with no effect on latency reactivation. IMPORTANCE Direct binding of HSV-1 ICP22 to the CD80 promoter downregulates the expression of the costimulatory molecule CD80 but not CD86. In this study, we fine mapped the region of ICP22 required for binding to the CD80 promoter and constructed a recombinant virus containing a deletion in ICP22 that failed to bind to the CD80 promoter. This recombinant virus replicated less efficiently in vitro and in vivo than did the WT control virus, although CD80-expressing CD11c+ cells and IFN-γ-expressing CD8+ T cells were increased. Interestingly, the levels of latency and reactivation in the two viruses were similar despite lower virus replication in the eyes of infected mice. Therefore, blocking the interaction of ICP22 with the CD80 promoter could be used to temper the immune response.

Ocular transmissibility of COVID-19: possibilities and perspectives.

Since the initial outbreak of coronavirus disease 2019 (COVID-19), extensive research has emerged from across the globe to understand the pathophysiology of this novel coronavirus. Transmission of this virus is a subject of particular interest as researchers work to understand which protective and preventative measures are most effective. Despite the well understood model of aerosol-respiratory mediated transmission, the exact mechanism underlying the inoculation, infection and spread of COVID-19 is currently unknown. Given anatomical positioning and near constant exposure to aerosolized pathogens, the eye may be a possible gateway for COVID-19 infection. This critical review explores the possibility of an ocular-systemic or ocular-nasal-pulmonic pathway of COVID-19 infection and includes novel insights into the possible immunological mechanisms leading to cytokine surge.

SARS-COV-2 viral load in tears of patients with COVID-19 in the early symptomatic stages: comparison of two different tear sampling methods.

PURPOSE: To evaluate the presence of SARS-CoV-2 virus in tears of patients with COVID-19 in the early symptomatic stages and to compare two different sampling methods. MATERIALS AND METHOD: In this cross-sectional study, tears sampling was performed in COVID-19 patients admitted within the first 7 days of symptom onset. The samples were collected with both conjunctival swabs and Schirmer strips. Each specimen was analyzed via RT-PCR. The viral load was evaluated in terms of the cycle threshold value. Ocular and systemic symptoms and comorbidities of the patients were also recorded. RESULTS: Forty patients were included. The average time from the initiation of symptoms was 3.15 days. Unilateral conjunctivitis has been observed in 5% of patients and foreign body sensation in 7.5% of patients. No viral RNA was detected in the tear samples of the patients with ocular findings. The positivity rate for SARS-CoV-2 in tears was 2.5% (n = 1). None of the samples collected by Schirmer test strips yielded positive polymerase chain reaction result for SARS-COV-2. The Ct value of the positive conjunctival swab was 36.03 and the nasopharyngeal Ct value of the same patient was 25.68. CONCLUSION: The SARS-CoV-2 viral shedding rate has been determined as 2.5% in the tears of early symptomatic stage COVID-19 patients. The viral load of the tears was lower than the naso-oropharynx. The conjunctival swab method is recommended in tear collection to evaluate the presence of SARS-CoV-2 by RT-PCR analysis in low viral load tears.

Evaluation of SARS-CoV-2 in Tears of Patients with Moderate to Severe COVID-19.

PURPOSE: To investigate the presence of SARS-CoV-2 RNA in tears of patients with moderate to severe coronavirus disease 2019 (COVID-19). DESIGN: Cross-sectional study. PARTICIPANTS: Patients with laboratory-proven moderate to severe COVID-19. METHODS: Tears were collected within 48 hours of laboratory confirmation using 3 methods: conjunctival swab plus Schirmer's test strips (group 1), conjunctival swab (group 2), and Schirmer's test strips (group 3). Samples from both the eyes of each patient were transported in a single viral transport media for real-time RT-PCR. Detailed demographic profiles, systemic symptoms, comorbidities, and ocular manifestations were noted. MAIN OUTCOME MEASURES: Viral load of a sample was determined using cycle threshold (Ct) value of E gene. A specimen was considered to show positive results if the amplification curve for the E gene crossed the threshold line within 35 cycles and if it showed positive results on an RNA-dependent RNA polymerase or open reading frame 1b gene assay. RESULTS: Of the 78 patients enrolled in the study, samples from 3 patients were found to be inadequate for analysis. Thirty-six patients (48%) had moderate disease, whereas 39 patients (52%) had severe disease, with no ocular involvement in any patient. In the 75 patients, RT-PCR analysis of tears showed positive results in 18 patients (24%), and 29 of 225 samples (12.9%) showed positive results. Positive results were found in 11 (14.7%), 11 (14.7%), and 7 (9.3%) patients in groups 1, 2, and 3, respectively (P = 0.3105). Mean Ct values in groups 1, 2, and 3 were 28.36 ± 6.15, 29.00 ± 5.58, and 27.86 ± 6.46 (P = 0.92), respectively. Five patients showed positive RT-PCR results by all 3 methods (mean Ct value, 25.24 ± 6.33), and 12 patients showed positive results by any of the 3 methods (mean Ct value, 32.16 ± 1.94), the difference in Ct values being statistically significant (P = 0.029). The median value of symptomatology in patients with positive RT-PCR results from tears was 5 days (range, 4-9 days). CONCLUSIONS: SARS-CoV-2 RNA was detected in tears of 24% of patients with laboratory-proven moderate to severe COVID-19. Conjunctival swab remains the gold standard of tear collection for RT-PCR assay. A significantly higher possibility of viral transmission exists through tears in patients with moderate to severe COVID-19.

An Eye on COVID-19: A Meta-analysis of Positive Conjunctival Reverse Transcriptase-Polymerase Chain Reaction and SARS-CoV-2 Conjunctivitis Prevalence.

SIGNIFICANCE: This analysis and review demonstrate that, although emerging data indicate that the prevalence of severe acute respiratory coronavirus 2 (SARS-CoV-2) on the ocular surface and coronavirus disease 2019 (COVID-19) conjunctivitis is rare, the ocular surface remains of interest as a potential inoculation and transmission site for SARS-CoV-2. Continued safety precautions should be taken as more data become available.COVID-19, caused by SARS-CoV-2, is a novel, global pandemic that has infected millions and, up to this point, caused more than two million fatalities worldwide. The ocular surface has become of interest as a possible vector for transmission by acting as a direct inoculation site, being a conduit for the virus into the respiratory system or as a method of transmission from potentially infected conjunctiva or tears. The components necessary for SARS-CoV-2 to theoretically infect ocular tissues are present: binding receptors (angiotensin-converting enzyme 2 and cluster of differentiation 147) and mechanisms for cell entry (transmembrane protease serine 2 and cathepsin L). This meta-analysis of COVID-19 prevalence data indicates that SARS-CoV-2 RNA has been infrequently found in conjunctival samples when tested with reverse transcriptase-polymerase chain reaction. This review estimates the prevalence of SARS-CoV-2 on the ocular surface and prevalence of conjunctivitis in patients with laboratory-confirmed COVID-19. There is much to be learned regarding ocular tropism of SARS-CoV-2.

Ocular Surface and Conjunctival Cytology Findings in Patients With Confirmed COVID-19.

PURPOSE: To assess the effect of severe acute respiratory syndrome coronavirus-2 infection on the conjunctiva and tear film. METHODS: Thirty-eight patients with confirmed COVID-19 and 31 healthy controls were included in this prospective and observational study. Individuals with COVID-19 formed the patient group, and healthy individuals formed the control group. Conjunctival impression cytology (CIC), TBUT, Schirmer II test, and ocular surface disease index were evaluated in all participants. RESULTS: No significant difference was observed regarding the mean age and gender between the groups (P=0.786 and P=0.122, respectively). The mean TBUT and Schirmer II test results did not differ between the two groups (P=0.496 and P=0.447, respectively). The CIC results revealed decreased density and cell size of goblet cells and moderate to high enlargement, squamous changes, and increased nucleocytoplasmic ratio in nongoblet epithelial cells in the COVID-19 group compared with the control group. Based on the Nelson classification in CIC samples, 60.6% of the COVID-19 group and 19.4% of the control group had changes consistent with grade 2 or above. The presence of neutrophils in CIC was significantly higher in the COVID-19 group (P<0.001), whereas the presence of lymphocyte was similar between the two groups (P=0.247). CONCLUSION: This study revealed the pathological conjunctival alterations in patients with COVID-19 and demonstrated that pathological ocular surface alterations may present even at the beginning of COVID-19 without clinically significant ocular manifestation.

Evaluation of coronavirus in tears and conjunctival secretions of patients with SARS-CoV-2 infection.

OBJECTIVE: This study aimed to assess the presence of novel coronavirus in tears and conjunctival secretions of SARS-CoV-2-infected patients. METHODS: A prospective interventional case series study was performed, and 30 confirmed novel coronavirus pneumonia (NCP) patients were selected at the First Affiliated Hospital of Zhejiang University from 26 January 2020 to 9 February 2020. At an interval of 2 to 3 days, tear and conjunctival secretions were collected twice with disposable sampling swabs for reverse-transcription polymerase chain reaction (RT-PCR) assay. RESULTS: Twenty-one common-type and nine severe-type NCP patients were enrolled. Two samples of tear and conjunctival secretions were obtained from the only one patient with conjunctivitis yielded positive RT-PCR results. Fifty-eight samples from other patents were all negative. CONCLUSION: We speculate that SARS-CoV-2 may be detected in the tears and conjunctival secretions in NCP patients with conjunctivitis.

Detection of HIV-1 viral load in tears of HIV/AIDS patients.

OBJECTIVES: The tear, as an important bodily secretion, plays a crucial role in preventing infection and maintaining homeostasis of ocular surfaces. Although accumulating studies have reported on the HIV-1 viral load profile among varying bodily fluids and secretions, little was known concerning HIV-1 dynamics in tears. Therefore, the objectives of this study were to investigate the HIV-1 viral load in tears of HIV/AIDS patients and study factors influencing their tear viral load. METHODS: A cross-sectional study was conducted. 67 patients with a confirmed HIV-1 infection or AIDS were recruited from the Beijing You'an Hospital, China between April 2018 and September 2018. Socio-demographic information and laboratory test results were collected. At the same time, ophthalmic examinations were carried out and tear samples were tested. RESULTS: Of 30 highly active antiretroviral therapy (HAART)-naïve patients, 53.3% had detectable HIV-1 RNA in tears. Of 37 patients on HAART, HIV-1 RNA was undetectable in their tears, regardless of treatment duration and blood viral load. Tear viral load ranged from TND (target not detected) to 13,096 copies/mL. Viral load was lower in tears than in blood plasma (p < 0.001), and was significantly correlated with plasma viral load (Rho = 0.566, p < 0.001) and AIDS stage (Rho = 0.312, p = 0.01), but negatively correlated with CD4 + T cell count, CD4 +/CD8 + T cell count, and duration of HIV infection (Rho = -0.450, Rho = - 0.464, Rho = - 0.565; p < 0.001). CONCLUSIONS: HIV-1 RNA is present in tears of more than half of the HAART-naïve patients, whereas absent in tears of patients on HAART. Tear viral load is positively associated with plasma viral load while it is negatively correlated with CD4 cell count. This study provides novel insights into the area with limited understanding-HIV-1 viral load in tears.

Ophthalmic manifestations of coronavirus disease 2019 and ocular side effects of investigational pharmacologic agents.

PURPOSE OF REVIEW: To compile and report the ocular manifestations of coronavirus disease 2019 (COVID-19) infection and summarize the ocular side effects of investigational treatments of this disease. RECENT FINDINGS: Conjunctivitis is by far the most common ocular manifestation of COVID-19 with viral particles being isolated from tears/secretions of infected individuals. Multiple therapeutic options are being explored across a variety of medication classes with diverse ocular side effects. SUMMARY: Eye care professionals must exercise caution, as conjunctivitis may be the presenting or sole finding of an active COVID-19 infection. While no currently studied therapeutic agents have been found to reliably treat COVID-19, early vaccination trials are progressing and show promise. A video abstract is available for a more detailed summary. VIDEO ABSTRACT: http://links.lww.com/COOP/A36.

COVID-19 pandemic from an ophthalmology point of view.

Coronavirus disease 2019 (COVID-19) is caused by a highly contagious RNA virus termed as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Ophthalmologists are at high-risk due to their proximity and short working distance at the time of slit-lamp examination. Eye care professionals can be caught unaware because conjunctivitis may be one of the first signs of COVID-19 at presentation, even precluding the emergence of additional symptoms such as dry cough and anosmia. Breath and eye shields as well as N95 masks, should be worn while examining patients with fever, breathlessness, or any history of international travel or travel from any hotspot besides maintaining hand hygiene. All elective surgeries need to be deferred. Adults or children with sudden-onset painful or painless visual loss, or sudden-onset squint, or sudden-onset floaters or severe lid oedema need a referral for urgent care. Patients should be told to discontinue contact lens wear if they have any symptoms of COVID-19. Cornea retrieval should be avoided in confirmed cases and suspects, and long-term preservation medium for storage of corneas should be encouraged. Retinal screening is unnecessary for coronavirus patients taking chloroquine or hydroxychloroquine as the probability of toxic damage to the retina is less due to short-duration of drug therapy. Tele-ophthalmology and artificial intelligence should be preferred for increasing doctor-patient interaction.

Ocular and Lacrimal Gland Lesions in Naturally Occurring Rabies of Domestic and Wild Mammals.

Investigations describing the ocular and lacrimal gland lesions associated with rabies are sparse. Here we characterize the pathological changes and distribution of rabies viral antigen in the eye, optic nerve, and lacrimal gland of 18 rabies cases from different mammalian species. Histology and immunohistochemistry for rabies virus, CD3, CD20, and Iba1 were performed on tissue sections of eye, optic nerve, and lacrimal gland. Polymerase chain reaction (PCR) for rabies was performed on all cases, including 7 formalin-fixed, paraffin-embedded (FFPE) and 11 frozen tissue samples of eye and lacrimal gland. Pathological changes in the eye consisted of retinal necrosis (12/18 cases) with occasional viral inclusions within ganglion cells (8/12 cases). Immunohistochemically, viral antigen was detected within the nerve fiber layer, ganglion cells, and inner plexiform layer in all 12 cases with retinal lesions and in 2 cases with no retinal lesions, as well as optic nerve (6/18 cases) and lacrimal gland epithelium (3/18 cases). CD3+ T lymphocytes were present in the retina (11/18 cases), optic nerve (2/18 cases), and lacrimal gland (11/18 cases). No CD20+ B lymphocytes or Iba1+ macrophages were detected. PCR for rabies virus was positive in 9 of 11 frozen samples but in only 2 of 7 FFPE samples. Five samples that were negative for rabies by PCR were positive by immunohistochemistry, and 2 samples were negative by both tests. These results provide evidence that rabies virus infection extends to the eye, likely via the ocular nerve, and that the lacrimal gland might be a source of viral infection.

There is a Role in Detection of SARS-CoV-2 in Conjunctiva and Tears: a comprehensive review.

Data on the involvement of the ocular surface and its relationship with Coronavirus disease 2019 (COVID- 19) are still minimal and not univocal. The respiratory tract is the structure most affected by COVID-19, and the serious form of the disease is characterized by severe pneumonia, acute respiratory distress syndrome and hypercoagulation. However, accumulating evidence shows that other organs could be reached by the virus, thus causing further comorbidities. To date, the exact route/routes of transmission of COVID-19 are still unclear. The respiratory tract is probably not the only route of transmission for this viral infection and some authors have also speculated that COVID-19 droplets, or infected hands, could contaminate the conjunctiva, which could therefore represent the initial site of an infection spread. Theoretically, the role of the ocular surface, a biological site still relatively unexplored, appears scientifically relevant in understanding the Severe Acute Respiratory Syndrome - Coronavirus - 2 (SARS-CoV-2) infection. The purpose of this paper is to summarize the current literature in order to elucidate the potential role of tear and conjunctival sampling to detect SARS-CoV-2 for the diagnosis of COVID-19 and to monitor patients during follow-up.

COVID-19: Ophthalmological Aspects of the SARS-CoV 2 Global Pandemic. / COVID-19: ophthalmologische Aspekte der globalen SARS-CoV-2-Pandemie.

PURPOSE: To perform a systematic analysis of articles on the ophthalmological implications of the global COVID-19 pandemic. METHODS: PubMed.gov was searched for relevant articles using the keywords "COVID-19", "coronavirus", and "SARS-CoV-2" in conjunction with "ophthalmology" and "eye". Moreover, official recommendations of ophthalmological societies were systematically reviewed, with a focus on the American Academy of Ophthalmology (AAO) and the Royal College of Ophthalmologists (RCOphth). RESULTS: As of April 16, 2020, in total, 21 peer-reviewed articles on the ophthalmological aspects of COVID-19 were identified. Of these, 12 (57.1%) were from Asia, 6 (28.6%) from the United States of America, and 3 (14.3%) from Europe. There were 5 (23.8%) original studies, 10 (47.6%) letters, 3 (14.2%) case reports, and 3 (14.2%) reviews. These articles could be classified into the topics "Modes and prevention of (ocular) transmission", "Ophthalmological manifestations of COVID-19", "Clinical guidance concerning ophthalmological practice during the COVID-19 pandemic", and "Practical recommendations for clinical infrastructure". Practical recommendations could be extracted from official statements of the AAO and the RCOphth. CONCLUSION: Within a short period, a growing body of articles has started to elucidate the ophthalmological implications of COVID-19. As the eye can represent a route of infection (actively via tears and passively via the nasoacrimal duct), ophthalmological care has to undergo substantial modifications during this pandemic. In the eye, COVID-19 can manifest as keratoconjunctivitis.

[A special on epidemic prevention and control: ophthalmologic research and prevention of 2019 novel coronavirus based on ocular manifestations of viral diseases].

This article was published ahead of print on the official website of Chinese Journal of Ophthalmology on February 24, 2020. In China, the fight against the 2019 novel coronavirus (2019-nCoV) has been at a critical stage. It has been confirmed that the transmission of 2019-nCoV is mainly through respiratory droplets and contact. Some scholars also pointed out that the possibility of transmission through the digestive system and eyes should not be ignored. Whether infection with 2019-nCoV will develop eye symptoms and whether the virus will spread through eyes are confusing to the medical workers and the general public, and it is ophthalmologists' responsibility to carry out in-depth discussions. Based on the ocular manifestations of viral diseases, this article analyzes whether the eye secretions and tears carry the virus, and whether ophthalmologists and patients are at a high risk for 2019-nCoV infection, and then presents the current research methods and the necessary prevention and control measures in the field of ophthalmology, with an aim to contribute to the fight against 2019-nCoV. ( Chin J Ophthalmol, 2020, 56: 414-417).

Evaluation of adenovirus amplified detection of immunochromatographic test using tears including conjunctival exudate in patients with adenoviral keratoconjunctivitis.

BACKGROUND: We evaluated a novel silver amplification immunochromatography test for rapid detection of adenovirus (AdV) antigen equipped with an automated reader system using tears including conjunctival exudate in patients with adenoviral keratoconjunctivitis. METHODS: Two kinds of immunochromatographic (IC) kits, a conventional IC kit for conjunctival scrapings (control kit) and an IC kit using tears including conjunctival exudate collected by pressing a filter paper strip on the conjunctiva (test kit), were tested on 90 patients who attended Migita Eye Clinic with suspected adenoviral conjunctivitis. The results of the test kits were automatically obtained by a specific reader, which was based on silver amplification immunochromatography system, in 15 min. The detection of AdV was confirmed by real-time polymerase chain reaction (PCR) method, and typing was performed by direct sequencing. Comparative dilution assay was carried out with the two kits, using AdV type 3 and type 54 strains. RESULTS: The sensitivity of the control kit and test kit was 89.8% and 98.3%, respectively. The specificity of both kits was 100%. A significant difference in the sensitivities of the two IC kits against PCR positivity was observed (P < 0.01). A significant correlation was found between AdV DNA copy numbers on a logarithmic scale obtained with the two tests (P < 0.01). The sensitivity of the test kit was 32-64-fold higher than that of the control kit without silver amplification for both AdV types. CONCLUSIONS: These results suggest that this novel amplified AdV detection kit using tears including conjunctival exudate is useful, because it decreases patients' discomfort from specimen collection and its sensitivity is significantly higher than that of the conventional IC kit.

Diagnostic tests for human rabies. / Tests diagnostiques de la rage chez l'homme.

The lack of reliable data concerning the number of human deaths from rabies presents one of the principal difficulties in a realistic assessment of the importance of this disease, and this lack of an accurate assessment has led to its underestimation and neglect. Priority should therefore be given to establishing a diagnostic test that can confirm human rabies on the basis of biological results. Indeed, only a laboratory diagnosis can properly identify infection, because clinical diagnosis remains difficult to interpret and is insufficiently specific. Historically, diagnosis has been based solely on post-mortem analysis of a cerebral biopsy using immunofluorescence techniques. Although this remains the standard method, considerable progress has been made with the advent of new molecular techniques and the evaluation of new, less-invasive sampling methods that are more easily accepted by the patient's family. Intra-vitam diagnosis of human rabies is now possible using reliable, robust, validated techniques that can be used everywhere, including in regions with limited resources, using minimally invasive or non-invasive sampling (such as saliva or skin biopsies). In practice, one of the major challenges with the diagnosis of human rabies is still the transfer and accessibility of such validated techniques in centralised reference laboratories located in low-income enzootic countries, in order to achieve the biological confirmation of each suspected case of rabies. At the same time, it is necessary to develop easy, fast and low-cost diagnostic methods that can be used in rural and remote areas in peripheral laboratories, or ideally at the patient's bedside.

L'absence de données fiables concernant le nombre de décès humains dus à la rage représente l'une des limitations majeures à l'évaluation réelle du poids mondial de cette maladie, contribuant ainsi à sa sous-estimation et à son caractère négligé. Devant ce constat, l'établissement d'un diagnostic de confirmation de la rage chez l'homme basé sur des résultats biologiques doit être favorisé. En effet, seul le diagnostic de laboratoire permet de valider l'infection, le diagnostic clinique restant difficile d'interprétation et insuffisamment spécifique. Historiquement, ce diagnostic était réalisé exclusivement au stade post-mortem via l'analyse d'une biopsie cérébrale par technique d'immunofluorescence. Bien qu'il s'agisse encore de la méthode de référence, des progrès considérables ont été faits, avec l'avènement de nouvelles techniques moléculaires et l'évaluation de nouveaux types de prélèvements moins invasifs et facilement acceptés par les proches du patient. Ces progrès autorisent maintenant la mise en oeuvre d'un diagnostic intra-vitam de la rage chez l'homme basé sur des techniques fiables, robustes et validées et pouvant être utilisées à tout niveau y compris dans les zones à ressources limitées à partir de prélèvements peu ou non invasifs (tels la salive ou les biopsies de peau). En effet, l'un des enjeux majeurs du diagnostic de la rage chez l'homme réside aussi dans le transfert et l'accessibilité de ces techniques validées, au niveau des laboratoires de référence situés dans les pays enzootiques à faible revenu, afin de réaliser une confirmation biologique de chaque cas suspect de rage. En parallèle, il est nécessaire de poursuivre les recherches sur le développement de méthodes de diagnostic simplifiées, rapides et de faible coût pouvant être utilisées de façon délocalisée, dans les laboratoires périphériques en zone rurale, voire au lit du patient.

La ausencia de datos fidedignos sobre el número de personas fallecidas a causa de la rabia constituye una de las principales limitaciones a la hora de evaluar con exactitud la carga mundial que impone la enfermedad, lo que contribuye al hecho de que esté subestimada y, por consiguiente, desatendida. De semejante constatación se desprende la necesidad de favorecer la instauración de un diagnóstico de confirmación de la rabia humana basado en resultados biológicos, en la medida en que el diagnóstico de laboratorio es el único modo de validar la presencia de la infección, pues el diagnóstico clínico presenta dificultades de interpretación y no es lo bastante específico. Históricamente este diagnóstico se realizaba únicamente tras la muerte del individuo, mediante el análisis por inmunofluorescencia de una muestra encefálica. Aunque este sigue siendo el método de referencia, el advenimiento de nuevas técnicas moleculares y el estudio de nuevos tipos de muestras, obtenidas por métodos menos invasivos y fácilmente aceptados por los allegados del paciente, han deparado progresos considerables, que permiten hoy realizar un diagnóstico intra-vitam de la rabia humana utilizando técnicas fiables, robustas y validadas que se pueden aplicar en todos los niveles, incluso en zonas con escasos recursos, a partir de muestras obtenidas por procedimientos poco o nada invasivos (muestras de saliva o biopsias de piel). Uno de los principales envites del diagnóstico de la rabia en el ser humano reside, en efecto, en la accesibilidad y la transferencia de estas técnicas validadas a laboratorios de referencia situados en los países enzoóticos de renta baja para poder realizar en ellos una confirmación biológica de todo caso sospechoso de rabia. Paralelamente, es necesario seguir investigando para instituir métodos de diagnóstico simplificados, rápidos y poco costosos que se puedan aplicar de forma descentralizada, esto es, en los laboratorios periféricos de zonas rurales e incluso junto al lecho del paciente.

The evaluation of diagnostic efficiency for stromal herpes simplex keratitis by the combination of tear HSV-sIgA and HSV-DNA.

PURPOSE: To assess the differential diagnostic values for stromal herpes simplex keratitis (HSK) by using tear HSV-sIgA, tear HSV-DNA, and the combination. METHODS: Tear samples for both eyes and the paired serum were collected from 187 stromal HSK and 56 controls. Enzyme-linked immune sorbent assay (ELISA) was used to analyze the tear HSV-sIgA and serum IgG/IgM/IgA. The levels of tear HSV-DNA were measured by polymerase chain reaction (PCR). RESULTS: The positive rates for tear HSV-sIgA and HSV-DNA were 36.90% and 10.96% respectively in stromal HSK patients. Twelve showed positivity for both sIgA and DNA, while 46 cases were positive for sIgA or DNA. The sensitivity, specificity, PPV, and NPV for simultaneous measurement were 39.73%, 98.21%, 98.31%, and 38.46%. The total negative conversion rate of sIgA was 95.71%. CONCLUSIONS: The diagnostic efficiency of HSV-sIgA only is nearly equal to the combination of HSV-sIgA and HSV-DNA, and the positive result is optimum to achieve a reliable diagnosis of stromal HSK even in atypical or unsuspected cases.

Herpes Simplex Virus Type 1 Shedding in Tears and Nasal and Oral Mucosa of Healthy Adults.

BACKGROUND: Herpes simplex virus type 1 (HSV-1) is prevalent worldwide and causes mucocutaneous infections of the oral area. We aimed to define the frequency and anatomic distribution of HSV-1 reactivation in the facial area in persons with a history of oral herpes. METHODS: Eight immunocompetent HSV-1 seropositive adults were evaluated for shedding of HSV-1 from 12 separate orofacial sites (8 from oral mucosa, 2 from nose, and 2 from conjunctiva) 5 days a week and from the oral cavity 7 days a week for approximately 5 consecutive weeks by a HSV DNA PCR assay. Symptoms and lesions were recorded by participants. RESULTS: Herpes simplex virus type 1 was detected at least from 1 site on 77 (26.5%) of 291 days. The most frequent site of shedding was the oral mucosa, with widespread shedding throughout the oral cavity. Lesional shedding rate was 36.4% (4 of 11 days with lesions), and the asymptomatic rate was 27.1% (65 of 240 nonlesional days). In individual participants, the median rate of HSV shedding by HSV PCR was 19.7% of days (range, 11%-63%). CONCLUSIONS: Reactivation of HSV-1 on the oral mucosa is common and usually asymptomatic. However, HSV-1 is rarely found in tears and nasal mucosa. Frequent oral shedding of HSV-1 may increase the risk for transmitting the virus to both oral and genital mucosa of sexual partners.

[Clinical presentations of Herpes Zoster Ophthalmicus (diagnosis and therapy)]. / Klinicheskie proyavleniya Herpes Zoster Ophthalmicus (diagnostika i terapiya).

Approximately a quarter of the world's population at some point in life is at risk of developing shingles (Herpes Zoster). In 10-20% of cases the first branch of the trigeminal nerve gets involved (Herpes Zoster Ophthalmicus, HZO). Ophthalmic complications of HZO are able to cause a significant reduction in visual function. AIM: To study and summarize clinical features of HZO (including the rate of complications and their nature) and to determine the relationship between clinical and laboratory data from these patients. MATERIAL AND METHODS: The study included 133 patients with ophthalmic and neurological complications of HZO (group 1 (n=28) - retrospective analysis of outpatient records for the period 1995-2005; group 2 (n=95) - a prospective study for the period 2005-2015), who received a course of conservative treatment in either the Botkin City Hospital, branch № 1, or in the ophthalmic department of the Moscow herpes centre (Gerpeticheskiy Tsentr Ltd.). Laboratory tests were performed only in patients from group 2 and included: examination of biological fluids for six types of herpes viruses by polymerase chain reaction, examination of tears and urine for DNA of Chlamydia, Mycoplasma, and Ureaplasma, and serological blood testing for markers of herpes virus infection. Patients from group 1 were prescribed topical antiviral, antibacterial, and anti-inflammatory therapy, in rare cases - acyclovir per os. In group 2, the treatment included systemic antiviral medications and immune correction therapy. Anti-inflammatory therapy consisted of local and systemic non-steroidal agents (NSAIDs). RESULTS: The most common ophthalmic complications of HZO in both groups were stromal keratitis and keratoiridocyclitis, neurological - III and VI cranial nerves palsies. The duration of the disease in the first group ranged from 2 months to 3 years; in the second group, patients were divided into two subgroups: subgroup A with the disease duration of no more than one month (n=81) and subgroup B with the disease duration from 1.5 to 9 months (n=14). Varicella-zoster virus (VZV) DNA was present in tears and/or other biological fluids of patients from group 2 in more than 70% of cases (n=67). Particularly, in 27.4% of cases the virus was isolated in two fluids and in 7.4% of cases - in three fluids. The duration of virus production in tears and other biological fluids (saliva, blood, and urine) ranged from 10 days to 4 months. CONCLUSION: Topical non-steroidal anti-inflammatory drugs and systemic etiological treatment in case of intraocular inflammation in HZO patients may reduce the risk of severe consequences of VZV reactivation and help avoid recurrences later in life.

Comparison of vaccine subpopulation selection, viral loads, vaccine virus persistence in trachea and cloaca, and mucosal antibody responses after vaccination with two different Arkansas Delmarva Poultry Industry -derived infectious bronchitis virus vaccines.

Factors responsible for the persistence of Arkansas Delmarva Poultry Industry (ArkDPI)-derived infectious bronchitis vaccines in commercial flocks and the high frequency of isolation of ArkDPI-type infectious bronchitis viruses in respiratory cases are still unclear. We compared dynamics of vaccine viral subpopulations, viral loads, persistence in trachea and cloaca, and the magnitude of infectious bronchitis virus (1BV)-specific antibody induction after vaccination with two commercial ArkDPI-derived Arkansas (Ark) serotype vaccines. One of the vaccines (coded vaccine B) produced significantly higher vaccine virus heterogeneity in vaccinated chickens than the other vaccine (coded A). Chickens vaccinated with vaccine B had significantly higher viral loads in tears at 5 days postvaccination (DPV) than those vaccinated with vaccine A. Vaccine B also induced a significantly higher lachrymal immunoglobulin M response at 11 DPV, an earlier peak of IBV-specific lachrymal immunoglobulin A, and higher serum antibodies than vaccine A. In addition, a significantly higher proportion of birds vaccinated with vaccine B had vaccine virus detected in the trachea at 20 DPV than those vaccinated with vaccine A. Furthermore, the virus detected at 20 DPV in most of the chickens vaccinated with vaccine B was a single specific subpopulation (subpopulation 4) selected from multiple vaccine subpopulations detected earlier at 5 and 7 DPV in the same chickens. On the other hand, a higher proportion of chickens vaccinated with vaccine A had virus detected in cloacal swabs at 20 DPV. Thus we found differences in mucosal antibody induction and selection and persistence of vaccine viruses between two ArkDPI-derived vaccines from different manufacturers. The higher vaccine virus heterogeneity observed in chickens vaccinated with vaccine B compared with those vaccinated with vaccine A may be responsible for these differences. Thus the high frequency of Ark IBV viruses in the field may be due to the inherent ability of some ArkDPI-derived vaccine viruses to be selected and persist in vaccinated chickens. Vaccine virus persistence may offer genetic material for recombination or may undergo mutations with the potential to result in increased virulence.