Paper Information

Journal:   JOURNAL OF CURRENT OPHTHALMOLOGY   2004 , Volume 17 , Number 1; Page(s) 30 To 36.
 
Paper: 

DIAGNOSIS OF ACANTHAMOEBA KERATITIS BY POLYMERASE CHAIN REACTION

 
 
Author(s):  KHOUSHZABAN F.*, DEYLAMIASL A.A.H., JABARVAND M., HASHEMIAN S.R., REZAEIAN M.
 
* Medical School of Shahed University
 
Abstract: 

Back Ground and Purpose: Some Acanthamoeba species can cause a chronic, progressive ulcerative keratitis, not responsive to antimicrobial therapy and frequently mistaken as stromal herpetic keratitis. In Acanthamoeba keratrtis (AK) many patients are initially treated with inappropriate drug therapies, hence, early detection of Acanthamoeba infection is critical. Contrary to molecular methods, use of parasite culturing as a diagnostic method may take several weeks. In the present work, PCR was used for diagnosing AK.
Materials and Methods: Corneal scraping specimens were collected from 70 patients referred to Farabi hospital as well as private clinics in Tehran. The specimens were cultured in ECNNA then the growing parasites were harvested. Phenol-chloroform method was applied for DNA extraction directly on corneal scraping specimens. Polymerase Chain Reaction was done for 18s rDNA amplification of the parasite using JDP1 and JDP2 as specific primers.
Results and Conclusion: Following PCR amplification of 18s rDNA of 70 specimens, 21 detected positive and a single band (458 bp) appeared on gel agarose and only 21 of the specimens (the same samples) grew in ECNNA culture after a long time. Therefore, PCR is fast and highly efficient in the diagnosis of Acanthamoeba keratitis.

 
Keyword(s): ACANTHAMOEBA, KERATITIS, PCR
 
References: 
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