title: Serological and polymerase chain reaction-based analysis of aqueous humour samples in patients with AIDS and necrotizing retinitis.
abstract: OBJECTIVE
  To evaluate the measurement of intraocular antibody production and detection of DNA by the polymerase chain reaction (PCR) for diagnosis of the causative microorganism in patients with AIDS and necrotizing retinitis.
  
  
  METHODS
  Paired serum and aqueous humour samples obtained from 28 patients with AIDS and necrotizing retinitis, seen between January 1987 and March 1992, were analysed for intraocular antibody production against cytomegalovirus (CMV), varicella zoster virus, herpes simplex virus, Epstein-Barr virus, and Toxoplasma gondii. Specific antibody titres in the inflamed eye and in the circulation were related to total immunoglobulin G content in the aqueous humour and serum. In addition, PCR analysis was performed in 15 samples. Results were compared to the final diagnosis, which was based on the subsequent clinical course. Results were also related to parameters describing the immune state of the patients: CD4 count, time between diagnosis of an AIDS-defining illness and retinitis, and time of survival following the diagnosis of retinitis.
  
  
  RESULTS
  In 11 (39%) out of 28 patients we found local intraocular antibody production which correlated with the final diagnosis (one out of two cases with acute retinal necrosis, three out of five cases with toxoplasma retinitis, and eight out of 21 patients with CMV retinitis). In all 13 patients with CMV retinitis PCR analysis detected CMV DNA. In one patient with the clinical diagnosis of Toxoplasma retinitis, Toxoplasma DNA could be determined, whereas in the same sample CMV DNA was also found. In yet another patient with Toxoplasma retinitis only CMV DNA could be detected. A relationship between results of local antibody determination with either CD4 counts, or the time interval between AIDS-defining illness and retinitis, or survival time after diagnosis of retinitis could not be established. CD4 counts were higher than 50 x 10(6)/l in eight out of 19 patients with CMV retinitis. No complications of paracentesis were seen.
  
  
  CONCLUSIONS
  Detection of intraocular antibody production and PCR analysis are quick and safe procedures and helpful tools for diagnosis of the involved pathogen in AIDS patients with a necrotizing retinitis. Negative results of local antibody production, even in the presence of detectable viral DNA, could not be related to the parameters of a more deteriorated immune status of these patients.
authors.affiliation: Department of Ophthalmology, Academic Medical Centre, University of Amsterdam, The Netherlands.
authors.forename: F D
authors.initials: FD
authors.lastname: Verbraak
authors.forename: M
authors.initials: M
authors.lastname: Galema
authors.forename: G H
authors.initials: GH
authors.lastname: van den Horn
authors.forename: M
authors.initials: M
authors.lastname: Bruinenberg
authors.forename: L
authors.initials: L
authors.lastname: Luyendijk
authors.forename: S A
authors.initials: SA
authors.lastname: Danner
authors.forename: A
authors.initials: A
authors.lastname: Kijlstra
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000150:complications*; D000328:Adult; D000914:Antibodies, Viral / Q000032:analysis; D001082:Aqueous Humor / Q000276:immunology / Q000469:parasitology* / Q000821:virology*; D017726:Cytomegalovirus Retinitis / Q000175:diagnosis; D003937:Diagnosis, Differential; D005260:Female; D005654:Fundus Oculi; D006801:Humans; D008297:Male; D008875:Middle Aged; D009336:Necrosis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012160:Retina / Q000473:pathology; D012173:Retinitis / Q000150:complications / Q000175:diagnosis*; D014123:Toxoplasmosis / Q000175:diagnosis
chemical_list: D000914:Antibodies, Viral