Determination the titer antibodies against LPS extracted from Pseudomonas aeruginosa isolated from eye infection

Abstract

rom September 2008 to May 2009, 122 specimens were collected by sterile cotton swaps from patients suffering from eye infection at Ibn Al-Haitham Teaching Eye Hospital, 12 of them (9.83% isolation percentage) were diagnosed as Pseudomonas aeruginosa isolates by using biochemical tests, API 20 E system: they were named (P(1), P(2),……..P(12)). The agglutination sera for the grouping of P. aeruginosa test show that all the isolates belong to the P. aeruginosa strains (6, 9, 12, 16). The result show that P10 isolate have a greater ability to adhesion when all P. aeruginosa isolates were tested on Congo red agar medium and adherence to smooth surfaces. The chemical analysis of crude LPS that was extracted from P10 by using digestive enzyme and hot phenol water method showed that the percentage of carbohydrates was 7.3 and the percentage of protein was 1.3. No nucleic acids were found while the chemical analysis of partially purified LPS that was made by gel-filtration chromatography by using Sephacryl 200 S showed that the percentage of carbohydrates in the partial purified LPS was 19.5% and the percentage of binding proteins was 0.006%. No nucleic acids were found. When the molecular weight of LPS was measured by gel-filtration chromatography by using Sepharose C1- 6B- 200 it was found to be 630957 Dalton. The sera were prepared by using two wild type rabbits (weight 2-2.5 kg). They were injected with five doses over 70 days of different concentrations of partially purified LPS that have been extracted from P. aeruginosa P (10) isolate. One rabbit was used as a control that has been injected with PBS. The titer of antibodies in sera was determined by passive haemagglutination and it was in the immunized rabbit’s sera 160, and the best concentration of LPS was 10µg.