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2.
Arq Bras Oftalmol ; 80(2): 84-87, 2017.
Article in English | MEDLINE | ID: mdl-28591279

ABSTRACT

OBJECTIVE:: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. METHODS:: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. RESULTS:: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. CONCLUSIONS:: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection.


Subject(s)
Cornea/virology , Herpesvirus 1, Human/isolation & purification , Herpesvirus 2, Human/isolation & purification , Herpesvirus 3, Human/isolation & purification , Keratitis, Dendritic/microbiology , Keratitis/microbiology , Real-Time Polymerase Chain Reaction/methods , Adolescent , Adult , Aged , Aged, 80 and over , Child , DNA Probes , Eye Infections, Bacterial/microbiology , Female , Humans , Keratitis/diagnosis , Keratitis/virology , Keratitis, Dendritic/diagnosis , Keratitis, Dendritic/virology , Male , Middle Aged , Prospective Studies , Young Adult
3.
Arq. bras. oftalmol ; 80(2): 84-87, Mar.-Apr. 2017. tab
Article in English | LILACS | ID: biblio-838786

ABSTRACT

ABSTRACT Objective: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. Methods: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. Results: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. Conclusions: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection.


RESUMO Objetivo: Ceratites bacterianas ocorrem mundialmente e apesar dos novos desenvolvimentos permanece como uma condição que pode levar à cegueira. Avaliar a presença de herpes simples (-1 e -2) e vírus varicella zoster (VZV) por reação em cadeia quantitativa de polimerase em tempo real (qPCR) em raspados corneanos de pacientes com ceratite bacteriana. Métodos: Sessenta e cinco pacientes com ceratite infecciosa foram submetidos a raspados corneanos estudados para gram, Giemsa, cultura e qPCR (grupo de estudo). Foram avaliados fatores de risco e epidemiológicos. O grupo controle foi composto por 25 casos de úlcera dendrítica típica por herpes analisados por qPCR. Resultados: Do grupo de estudo (n=65), nove pacientes (13,8%) apresentaram cultura, qPCR e raspado negativos. Cinquenta e seis (86,2%) pacientes apresentaram cultura positiva, 51 para bacteria, 4 para fungo e 1 para ameba. A qPCR identificou 10 pacientes do grupo de cultura positiva para bactéria que também foram positivos para vírus, um VZV e 9 para HSV-1. Dos 25 pacientes que compunham o grupo controle, 21 apresentaram qPCR positivo para HSV-1. Conclusão: Herpes pode estar presente em pacientes com úlceras de córnea bacterianas e a qPCR pode ser útil na sua detecção.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Keratitis, Dendritic/microbiology , Herpesvirus 2, Human/isolation & purification , Herpesvirus 1, Human/isolation & purification , Herpesvirus 3, Human/isolation & purification , Cornea/virology , Real-Time Polymerase Chain Reaction/methods , Keratitis/microbiology , DNA Probes , Eye Infections, Bacterial/microbiology , Keratitis, Dendritic/diagnosis , Keratitis, Dendritic/virology , Prospective Studies , Keratitis/diagnosis , Keratitis/virology
5.
Yan Ke Xue Bao ; 9(3): 126-8, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8168606

ABSTRACT

HSV-DNA of seven corneal lesions suspected with herpes simplex keratitis (HSK) and nine normal human donor corneas were detected by PCR. Five out of seven diseased corneas showed positive results, and the other two diseased corneas and nine normal corneas negative. The results suggest the PCR may be useful as a rapid and sensitive method for diagnosing HSK.


Subject(s)
DNA, Viral/analysis , Herpesvirus 1, Human/isolation & purification , Keratitis, Herpetic/microbiology , Adult , Base Sequence , Cornea/microbiology , Female , Herpesvirus 1, Human/genetics , Humans , Keratitis, Dendritic/microbiology , Male , Middle Aged , Molecular Sequence Data , Polymerase Chain Reaction
6.
Cornea ; 11(5): 471-4, 1992 Sep.
Article in English | MEDLINE | ID: mdl-1330439

ABSTRACT

Although varicella is one of the most common infectious diseases in the United States, systemic and ocular complications are rare. We report a patient who developed disciform edema followed by microdendritic keratitis 1 and 2 months, respectively, after resolution of the acute phase of varicella. Cultures were negative, but serologic analysis found positive antibodies against varicella zoster virus and negative antibodies against herpes simplex virus. Based on this case and on a review of the literature, we believe that this delayed onset of keratitis represents a distinct category of varicella corneal complications.


Subject(s)
Chickenpox , Keratitis, Dendritic/microbiology , Antibodies, Viral/analysis , Chickenpox/drug therapy , Child , Corneal Edema/drug therapy , Corneal Edema/microbiology , Female , Herpesvirus 3, Human/immunology , Humans , Keratitis, Dendritic/drug therapy , Prednisone/therapeutic use , Recurrence , Trifluridine/therapeutic use , Vidarabine/therapeutic use , Visual Acuity
8.
Ophthalmology ; 99(5): 781-99, 1992 May.
Article in English | MEDLINE | ID: mdl-1317538

ABSTRACT

The herpes simplex and varicella-zoster viruses are members of the subfamily alpha herpesviruses with specific properties of the virion and with the capacity to establish latent infections in humans. The genome of each of these viruses has been determined with an estimate of the number of genes and proteins encoded. The biology and molecular events of the herpes simplex virus productive and latent infection have been detailed with the use of both in vitro and in vivo model systems. The neuron is the site of latency in the ganglia with a limited transcription of genes expressed during the latent period. The specific molecular regulation of latency and reactivation are not well established. There are co-cultivation, electron microscopy, and biochemical studies that support the concept of corneal latency, although this has not been proven conclusively. Details about the varicella-zoster virus biology and molecular events are not as well advanced since animal models have been lacking. The biology of the productive infection (varicella) is different from herpes simplex virus infection since the portal of entry is the respiratory system. Data support the concept of the maintenance of latency within satellite cells in the ganglia rather than within neurons. There are multiple genes expressed during this latency. These features may explain the different clinical presentations and course of reactivation (zoster) compared with herpes simplex virus reactivation.


Subject(s)
Eye Infections, Viral/microbiology , Herpesvirus 3, Human/physiology , Keratitis, Dendritic/microbiology , Simplexvirus/physiology , Animals , Chickenpox/microbiology , Cornea/innervation , Ganglia/microbiology , Gene Expression Regulation, Viral , Genes, Viral , Herpesvirus 3, Human/genetics , Humans , Neurons/microbiology , Simplexvirus/genetics , Virus Activation/physiology , Virus Replication/physiology
10.
Invest Ophthalmol Vis Sci ; 32(10): 2741-6, 1991 Sep.
Article in English | MEDLINE | ID: mdl-1654309

ABSTRACT

The authors characterized a murine model of herpes simplex virus (HSV) reactivation in which recurrent herpetic keratitis was obtained in up to 80% of animals. Five weeks after ganglionic latency was established in National Institutes of Health inbred mice after corneal inoculation, HSV type 1 (HSV-1) was reactivated by irradiating the previously inoculated eye with ultraviolet (UV) light. Comparison of different UV wavelengths showed UVB to be optimal for reactivation, with peak viral recurrence being induced by a total exposure of approximately 250 mJ/cm2. Reactivated infectious virus generally began to appear in trigeminal ganglia 2 days postirradiation and was subsequently detectable in the cornea by both corneal swabbing and immunostaining for viral antigens. Two consecutive outbreaks of viral recurrence at the ocular surface were induced in selected animals by serial exposure to UVB. Advantages of this model over other models of recurrent keratitis are discussed.


Subject(s)
Keratitis, Dendritic/microbiology , Simplexvirus/growth & development , Ultraviolet Rays , Virus Activation/radiation effects , Animals , Antigens, Viral/immunology , Cornea/innervation , Cornea/microbiology , Cornea/radiation effects , Disease Models, Animal , Mice , Mice, Inbred Strains , Recurrence , Simplexvirus/immunology , Simplexvirus/isolation & purification , Simplexvirus/radiation effects , Trigeminal Ganglion/microbiology , Vero Cells
11.
J Gen Virol ; 72 ( Pt 9): 2043-9, 1991 Sep.
Article in English | MEDLINE | ID: mdl-1654368

ABSTRACT

We used a herpes simplex virus (HSV) type 1 ribonucleotide reductase (RR) null mutant (ICP6 delta) to study the role of HSV-1 RR in ocular HSV infections. We found that ICP6 delta was unable to induce vascularization of the cornea or stromal keratitis following inoculation into the cornea of BALB/c mice, but was able to induce a transient mild blepharitis. The parental strain (HSV-1 KOS) and a revertant of ICP6 delta, ICP6 delta+3.1, both caused severe ocular disease, indicating that HSV-1 RR is required for ocular virulence in mice. ICP6 delta grew poorly in vitro (Vero and BALB/c 3T3 fibroblasts) and in vivo (eye, trigeminal ganglia and brain) compared to ICP6 delta+3.1 and HSV-1 KOS, suggesting that the avirulence of ICP6 delta is due to poor growth in the host. ICP6 delta also grew less well in primary human corneal fibroblasts, suggesting that RR may be required for virulence in humans. These results indicate that drugs inhibiting the function of RR might be effective in treating ocular HSV infections.


Subject(s)
Blepharitis/microbiology , Keratitis, Dendritic/microbiology , Ribonucleotide Reductases/metabolism , Simplexvirus/enzymology , Animals , Brain/microbiology , Cell Line , Cornea/microbiology , Female , Fibroblasts/microbiology , Humans , Mice , Mice, Inbred BALB C , Mutagenesis , Ribonucleotide Reductases/genetics , Simplexvirus/genetics , Simplexvirus/pathogenicity , Vero Cells , Virulence
13.
J Virol ; 65(8): 4142-52, 1991 Aug.
Article in English | MEDLINE | ID: mdl-1649322

ABSTRACT

In a mouse model for herpes simplex virus type 1 (HSV-1) latency in which the virus was inoculated via the eye after corneal scarification, HSV-1 replicated in corneal epithelial cells and infected the nerve cell endings. HSV-1 reached the trigeminal ganglia by fast axonal transport between 2 and 10 days postinfection (p.i.) and established a latent infection in neuronal cells or replicated and spread to nonneuronal cells. By using in situ hybridization, we showed that cellular transcription factors are stimulated by HSV-1 infection in trigeminal ganglia. This stimulation is biphasic, peaking at 1 and 3 to 4 days p.i. The first peak involves c-jun and oct-1 expression in neurons, and the second involves c-jun, c-fos, and oct-1 expression in neurons and nonneuronal cells. Corneal scarification, alone or followed by infection with UV-inactivated HSV-1, induced monophasic c-jun and oct-1 expression in some neurons of the trigeminal ganglia, with a peak at 1 day p.i. Corneal infection without prior scarification induced c-jun, c-fos, and oct-1 expression in some neuronal and nonneuronal cells of the trigeminal ganglia 2 to 9 days p.i. Explanation of ganglia from latently infected animals resulted in reactivation of the latent virus. Independently of the presence of latent HSV-1 in explanted ganglia, expression of c-fos, c-jun, and oct-1 was induced first in nonneuronal cells, peaking 6 to 10 h postexplantation, and then in neuronal cells, with a peak at 24 h after explantation when expression of viral replicative genes was first detectable. Since ocular HSV-1 infection, corneal scarification, and explantation of trigeminal ganglia all resulted in induction of expression of cellular transcription factors in ganglia, these factors may play a critical role in the permissiveness of cells for HSV-1 replication during acute infection, latency, and reactivation.


Subject(s)
Keratitis, Dendritic/microbiology , Simplexvirus/physiology , Transcription Factors/biosynthesis , Trigeminal Ganglion/microbiology , Animals , Cornea/microbiology , DNA Probes , DNA, Viral/analysis , DNA-Binding Proteins/biosynthesis , Disease Models, Animal , Female , Host Cell Factor C1 , Mice , Mice, Inbred BALB C , Nucleic Acid Hybridization , Octamer Transcription Factor-1 , Organ Culture Techniques , Proto-Oncogene Proteins/biosynthesis , Proto-Oncogene Proteins c-fos , Proto-Oncogene Proteins c-jun , Simplexvirus/genetics , Transcription, Genetic , Virus Replication
14.
Invest Ophthalmol Vis Sci ; 32(6): 1808-15, 1991 May.
Article in English | MEDLINE | ID: mdl-1851732

ABSTRACT

Herpes simplex virus (HSV) latency in sensory ganglion neurons is well documented, but the existence of extraneuronal corneal latency is less well defined. To investigate the possibility of extraneuronal latency during ocular HSV infection, corneal specimens from 18 patients with quiescent herpes simplex keratitis (HSK) were obtained at the time of keratoplasty. Polymerase chain reaction (PCR) amplification followed by southern blot hybridization with a radiolabeled oligonucleotide probe was done to detect the presence of HSV-1 genome in these human corneal samples. Two pairs of oligonucleotides from the region of the HSV thymidine kinase (TK) gene and the latency-associated transcript (LAT) gene were used as primers in the PCR amplification. The DNA sequences from either the TK or the LAT gene were identified in 15 of 18 HSK corneas (83%). These results demonstrate that the HSV genome was retained, at least in part, in human corneas during quiescent HSV infection, giving further support to the concept of corneal extraneuronal latency.


Subject(s)
Keratitis, Dendritic/microbiology , Polymerase Chain Reaction , Simplexvirus/genetics , Thymidine Kinase/genetics , Adult , Aged , Aged, 80 and over , Base Sequence , Blotting, Southern , DNA, Viral/analysis , Female , Gene Amplification , Humans , Male , Middle Aged , Molecular Sequence Data , Oligonucleotide Probes , Simplexvirus/enzymology
15.
Invest Ophthalmol Vis Sci ; 32(6): 1816-20, 1991 May.
Article in English | MEDLINE | ID: mdl-1851733

ABSTRACT

The phosphonylmethoxyalkyl derivative, (S)-1-(3-hydroxy-2-phosphonyl methoxypropyl)cytosine (HPMPC), was evaluated for its efficacy in the topical treatment of experimental keratitis caused by thymidine kinase-positive (TK+) or thymidine kinase-deficient (TK-) herpes simplex virus type 1 (HSV-1) strains. The HPMPC 0.2% eyedrops were as effective as the reference compound, (E)-5-(2-bromovinyl)-2'-deoxyuridine, (BVDU) 0.2% eyedrops in stimulating the healing of epithelial disease caused by the HSV-1 TK+ strain. Both drugs achieved a significant (P less than 0.005) healing effect compared with placebo eyedrops. No significant differences were noted in the efficacy of HPMPC 0.2% eyedrops when instilled one, three, or nine times a day. In the treatment of keratitis caused by the HSV-1 TK- strain, 0.2% BVDU eyedrops were similar to placebo; 0.2% HPMPC eyedrops again had a brisk and significant healing effect (P less than 0.005).


Subject(s)
Antiviral Agents/therapeutic use , Cytosine/analogs & derivatives , Keratitis, Dendritic/drug therapy , Organophosphonates , Organophosphorus Compounds/therapeutic use , Simplexvirus/enzymology , Thymidine Kinase/metabolism , Animals , Antiviral Agents/administration & dosage , Bromodeoxyuridine/administration & dosage , Bromodeoxyuridine/analogs & derivatives , Bromodeoxyuridine/therapeutic use , Cidofovir , Cytosine/administration & dosage , Cytosine/therapeutic use , Disease Models, Animal , Drug Administration Schedule , Female , Keratitis, Dendritic/microbiology , Male , Ophthalmic Solutions , Organophosphorus Compounds/administration & dosage , Rabbits , Random Allocation
16.
Am J Ophthalmol ; 111(4): 480-4, 1991 Apr 15.
Article in English | MEDLINE | ID: mdl-1849354

ABSTRACT

We treated three patients with herpes simplex dendritic keratitis that occurred between three and 11 months after keratoplasty. The patients had no history of herpetic infection. The eyes of two of the patients were grafted for corneal scarring of undetermined origin. The eye of the third patient was grafted for pseudophakic bullous keratopathy. At the time of onset of dendritic keratitis, all three patients were receiving either maintenance or higher doses of topical corticosteroids. All infections responded to topical antiviral treatment. The findings in these patients illustrate the importance of considering herpes simplex keratitis in the differential diagnosis of all late-onset epithelial defects in the corneal graft, even in the absence of a history of herpes simplex keratitis.


Subject(s)
Keratitis, Dendritic/etiology , Keratoplasty, Penetrating/adverse effects , Administration, Topical , Adrenal Cortex Hormones/therapeutic use , Aged , Cornea/microbiology , Diagnosis, Differential , Fluorescent Antibody Technique , Humans , Keratitis, Dendritic/drug therapy , Keratitis, Dendritic/microbiology , Male , Middle Aged , Simplexvirus/isolation & purification , Trifluridine/therapeutic use , Visual Acuity
17.
Invest Ophthalmol Vis Sci ; 32(5): 1558-61, 1991 Apr.
Article in English | MEDLINE | ID: mdl-1849874

ABSTRACT

A mouse model for ocular reactivation of herpes simplex virus type 1 (HSV-1) was modified and used to study the effect of strain difference on the frequency of ocular HSV reactivation. Outbred male NIH white mice were immunized with 1.0 ml of anti-HSV serum with a neutralizing titer of 1:400 24 hr before infection and bilaterally infected at 10(5) plaque-forming units/eye with one of three HSV-1 strains: 17 Syn+, LAT+ (XC-20), or LAT- (X10-13). Latency-associated transcripts (LAT) are produced by strain 17 Syn+ and LAT+ but not by LAT-. The primary infection was monitored by ocular swabbing for HSV. Reactivation was induced by intravenous (i.v.) injection of cyclophosphamide (5 mg) followed 24 hr later by i.v. dexamethasone (0.2 mg). These drugs significantly reduced the white cell count between 0 and 6 days post-administration. The eyes were swabbed for 7 consecutive days to monitor reactivation, and HSV-1 reactivation was induced at the following frequencies in individual eyes: 17 Syn+ (32.5%), LAT+ (18.5%), and LAT- (2.5%) (P less than or equal to 0.002). Co-culture of trigeminal ganglia was done, and random isolates were checked to ascertain their identity. The HSV was recovered from individual trigeminal ganglia at the following frequencies: 17 Syn+ (83%), LAT+ (100%), and LAT- (67%) (P less than or equal to 0.091). These results confirm that the mouse can be used as a reactivation model for ocular HSV infection and that the presence of LAT facilitates reactivation in vivo in the mouse.


Subject(s)
Genes, Viral , Keratitis, Dendritic/microbiology , Simplexvirus/genetics , Virus Activation , Animals , Antibodies, Viral/administration & dosage , Cornea/microbiology , Cyclophosphamide/administration & dosage , Dexamethasone/administration & dosage , Disease Models, Animal , Keratitis, Dendritic/immunology , Male , Mice , Simplexvirus/growth & development , Simplexvirus/immunology , Trigeminal Ganglion/microbiology , Virus Activation/drug effects
19.
Br J Ophthalmol ; 75(4): 195-200, 1991 Apr.
Article in English | MEDLINE | ID: mdl-1850616

ABSTRACT

Patients undergoing penetrating keratoplasty for prior herpes simplex keratitis (group A) and corneal disease unrelated to herpes simplex (group B) were investigated to assess whether the cornea is a site for herpes simplex viral latency. All patients were seropositive for herpes simplex viral antibody. Virus was isolated from the tear film postoperatively in one patient and on cocultivation from the cornea of another patient. Herpes simplex viral DNA, however, was detected in the corneas of all patients from group A and half of those from group B by means of the polymerase chain reaction and primers to three well separated regions of the viral genome. Three donor corneas had no evidence of herpes simplex viral DNA. Using RNA polymerase chain reaction, we found evidence of a latency associated transcript and also that of a glycoprotein C coding transcript in two corneas, indicating viral replication. Nine corneas had evidence of a latency associated transcript but no glycoprotein C transcript, which suggests that herpes simplex virus may be maintained in a latent state in the corneas of patients with prior herpes simplex keratitis and in some patients with corneal disease unrelated to the herpes simplex virus.


Subject(s)
Cornea/microbiology , Keratitis, Dendritic/microbiology , Simplexvirus/isolation & purification , Adult , Aged , Base Sequence , Corneal Diseases/microbiology , Corneal Transplantation , DNA, Viral/analysis , Female , Humans , Male , Middle Aged , Molecular Sequence Data , Polymerase Chain Reaction , RNA, Viral/analysis , Simplexvirus/genetics , Tears/microbiology , Transcription, Genetic
20.
Zhonghua Yan Ke Za Zhi ; 27(2): 109-11, 1991 Mar.
Article in Chinese | MEDLINE | ID: mdl-1650312

ABSTRACT

The authors first report in China the detection of HSV-1 antigen in 36 human corneas with recurrent HSK, using the horse radish peroxidase staining technique. The results were that in 16 eyes of quiescent HSK, the HSV antigen was negative, and in 11 of 20 corneas with active HSK, the HSV antigen was positive. When 5 corneas with quiescent HSK were cultured in vitro, 3 again became HSV antigen positive. The findings may be useful in the treatment of HSK.


Subject(s)
Antigens, Viral/analysis , Cornea/immunology , Keratitis, Dendritic/immunology , Simplexvirus/immunology , Cornea/microbiology , Humans , Immunohistochemistry , Keratitis, Dendritic/microbiology , Recurrence
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