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Maria Valeria C Pereira Silva, Erna G Kroon, Fernando C Trindade, Mauricio L Nogueira, Eduardo A Bambirra; Detection of Human herpes virus 1 (HHV-1) in graft failure corneas: a study of 24 cases using polymerase chain reaction (PCR). Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1860.
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© ARVO (1962-2015); The Authors (2016-present)
The purpuse of this study was to evaluate the presence of Human herpes virus 1 (HHV-1) in paraffin-embedded corneas of graft failure using a polymerase chain reaction protocol.
Twenty four corneas obtained by penetratin keratoplasty (PK) were fixed in 70% alcohool and paraffin embedded. From each patient clinical data was collected such as gender, age and surgical indications.In cases of previous corneal graft failure, we collected information about previous PK indication, such as: how long the graft remained transparent, whether the patient had glaucomaand the time interval between the corneal graft failure and the new surgery.The samples were submitted to DNA extraction and PCR DNA amplification.Primers for HHV-1 thymidine kinase and interferon beta gene were used in PCR. Clinical data from 18 patients were analyzed. In six cases the clinical data was not available.
The male/female ratio was 50%. The age average of analyzed patients was 53.9 years with a standard deviation of 17.4 years. The corneal transplant indication occurred in 13 (72%) cases because of graft failure, the other 5 cases (27.5%) due to one of the following pathologies each: bullous kerathopathy, fungus keratitis, traumatic corneal opacity, bacterial ulcer and corneal opacity presumed to be secondary to herps keratitis happened six years before. The time period in which the graft remained clear in failure cases varied from immediate, primary failure to 34 months after surgery, with an average of 10.0 months. Seven patients (54%) had glaucoma diagnosis among patients that had a corneal graft because of graft failure. The time period between the graft failure and the new surgery varied from one to 132 months, with an average of 30.8 months with a standard deviation of 34.6 months. The developed protocol showed to be able to detect up to 0.001 plaque forming unit/ml of the HHV-1. By using this protocol, our samples were negative for HHV-1 DNA.
This technique may be of clinical importance in the study of HHV-1 influence in corneal graft failure.
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