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      Clinical spectrum, diagnostic criteria, and polymerase chain reaction of aqueous humor in viral and toxoplasma detection in Fuchs’ uveitis syndrome

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          Abstract

          Aim:

          The aim of this study is to describe the clinical features and diagnostic criteria of Fuchs’ uveitis (FU) and to determine whether it has an association with virus and toxoplasma in the aqueous humor during cataract surgery.

          Setting and Design:

          This is a prospective, case–control study.

          Materials and Methods:

          Patients with FU ( n = 25), anterior uveitis ( n = 15), and no uveitis (normal) ( n = 50) were included based on predefined inclusion and exclusion criteria for all three groups. Polymerase chain reaction (PCR) of aqueous humor and serum for rubella, herpes simplex virus (HSV), cytomegalovirus (CMV), varicella-zoster virus (VZV), and toxoplasma was done using conventional uniplex PCR.

          Statistical Analysis:

          It was done using SPSS software using Chi-square test for categorical variables, and P < 0.05 was considered statistically significant.

          Results:

          Ninety patients were enrolled in the study in three groups, comparable for age, gender, and laterality of ocular involvement. All patients had diffuse keratic precipitates in FU group ( P = 0001) with none having posterior synechiae ( P = 0.046) which was statistically significant when compared to anterior uveitis patients. Iris nodules were noted in one case in both groups. Serum and aqueous PCR was negative for detection of VZV, CMV, toxoplasma, and rubella in all groups. PCR for HSV was positive in one patient in “normal” group but was not statistically significant.

          Conclusion:

          Our study shows that diagnosis of FU is mainly clinical. There appears to be no role of aqueous humor testing for viruses by PCR to aid in etiological diagnosis.

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          Most cited references21

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          Fuchs heterochromic cyclitis: rubella virus antibodies and genome in aqueous humor.

          To characterize the polyspecific intraocular antibody synthesis in aqueous humor of patients with chronic inflammatory diseases of the eye and to detect the causative antigen in Fuchs heterochromic cyclitis (FHC). Retrospective case-control study. Intraocular antibody synthesis is detected in aqueous humor with the Antibody Index [AI] (improved Goldmann-Witmer Index) and quantified as specific antibody fraction, F(s) (intraocular specific antibody concentration in percent of intraocular total immunoglobulin G in aqueous humor). Virus detection is by nested polymerase chain reaction. Fifty-two eyes of 52 patients with clinically defined FHC (aged 16-73 years) had an intraocular synthesis of rubella antibodies (AI > or =1.5). The rubella genome was detected in 5 (18%) of 28 aqueous humor samples investigated, or in 5 (56%) of 9 patients aged <40 years. Oligoclonal IgG was synthesized in 34 (87%) of 39 eyes. Unaffected fellow eyes (n = 3) or cerebrospinal fluid (n = 2) were normal. In FHC the median rubella AI = 20.6 (total range 1.5-309) was seven times higher than in multiple sclerosis (MS) patients (n = 15) with uveitis intermedia or periphlebitis retinae. In MS the intraocular rubella antibody synthesis (frequency 73%) is part of a polyspecific immune response (increased measles AI in 80%, varicella zoster virus AI in 47%, herpes simplex virus AI in 23%). The median rubella-F(s) = 2.6% in FHC (range = 0.14%-45.9%) was approximately 40 times higher than in MS, consistent with a virus-driven antibody response in FHC. Noninflammatory controls (50 senile cataracts) had neither an intraocular rubella antibody synthesis (normal AI < or =1.4) nor rubella antigen in aqueous humor. The rubella AI was normal in all patients with an intraocular toxoplasmosis (n = 24), anterior uveitis (n = 27), herpes simplex virus iritis (n = 25), and varicella zoster virus iritis (n = 14). Fuchs heterochromic cyclitis is a rubella virus-driven disease with persistence of the virus preferentially detected in the younger patients. The proposed laboratory supported diagnosis of FHC is based on the increased rubella Antibody Index. The virus etiology gives a rationale for omitting the ineffective corticosteroid therapy of FHC.
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            Comparison of PCR detection methods for B1, P30, and 18S rDNA genes of T. gondii in aqueous humor.

            Comparison of polymerase chain reaction (PCR) amplification of three Toxoplasma gondii genes in aqueous humor. Nested PCRs carried out using published methods were optimized for maximum sensitivity and specificity. Five pairs of oligonucleotide primers, directed against the B1, P30, and ribosomal genes, were used and compared to determine which sequences were most effective in detecting T. gondii DNA. Methods were developed with DNA templates in water and were subsequently applied to both normal and inflamed aqueous. After one round of PCR amplification, P30 and ribosomal primers were able to detect 1 pg genomic T. gondii DNA. However, those directed against the B1 gene were able to detect 50 fg (approximately single tachyzoite). This level of sensitivity was also achieved using the P30 primers after a second round of PCR; however, only primers based on the B1 gene maintained this level of sensitivity in both normal and inflamed aqueous. B1-specific primers did not amplify sequences from fungal, bacterial, or human lymphocyte DNA. The sensitivity of T. gondii detection using B1 gene-specific primers was not compromised when large amounts of human lymphocyte DNA were present, and application to an ocular sample or retinal section from patients with toxoplasma chorioretinitis was successful in confirming the presence of T. gondii DNA. The B1 PCR protocol appears to be the most sensitive protocol in the detection of T. gondii DNA and has been successful in identification of T. gondii DNA in ocular fluids and retinal sections. This provides direct evidence of the presence of T. gondii within the eye and may therefore help in the management of toxoplasma retinochoroiditis.
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              Rubella virus is associated with fuchs heterochromic iridocyclitis.

              To determine whether rubella virus (RV) is involved in the pathogenesis of Fuchs heterochromic iridocyclitis (FHI). Retrospective patient-controlled study. Intraocular immunoglobulin G production against RV, herpes simplex virus (HSV), varicella zoster virus (VZV), and Toxoplasma gondii was determined in the aqueous humor of 14 patients with FHI, 13 control subjects with herpetic uveitis anterior, and 19 control subjects with ocular toxoplasmosis by calculation of the Goldmann-Witmer coefficient (GWC). All patients and control subjects were seropositive for RV. Intraocular antibody production (GWC >3) against RV was found in 13 of 14 patients (93%) with FHI. Intraocular antibody production against HSV, VZV, or T gondii was not detected. None of the control subjects with herpetic uveitis anterior or with toxoplasma chorioretinitis had a positive GWC for rubella virus (P < .0001, Fisher exact test). Rubella virus, but not HSV, VZV, or T gondii, is associated with FHI.
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                Author and article information

                Journal
                Indian J Ophthalmol
                Indian J Ophthalmol
                IJO
                Indian Journal of Ophthalmology
                Medknow Publications & Media Pvt Ltd (India )
                0301-4738
                1998-3689
                August 2016
                : 64
                : 8
                : 555-558
                Affiliations
                [1]Department of Cornea and Anterior Segment, Tej Kohli Cornea Institute, L.V. Prasad Eye Institute, Hyderabad, Telangana, India
                [1 ]Jhaveri Microbiology Center, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
                [2 ]Center for Ocular Regeneration, Srujana Innovation Center, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
                [3 ]Department of Clinical Research, L. V. Prasad Eye Institute, Hyderabad, Telangana, India
                Author notes
                Correspondence to: Dr. Somasheila I Murthy, Cornea and Anterior Segment Services and Uveitis Services, L. V. Prasad Eye Institute, Kallam Anji Reddy Campus, L. V. Prasad Marg, Banjara Hills, Hyderabad - 500 034, Telangana, India. E-mail: smurthy@ 123456lvpei.org
                Article
                IJO-64-555
                10.4103/0301-4738.191485
                5056540
                27688274
                dcafddda-ab32-4cab-9e9b-fe90403a1e8e
                Copyright: © Indian Journal of Ophthalmology

                This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

                History
                : 12 November 2015
                : 04 July 2016
                Categories
                Original Article

                Ophthalmology & Optometry
                fuchs’ uveitis,keratic precipitates,polymerase chain reaction,posterior synechiae

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