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Serum IgG response to an outer membrane porin protein of Burkholderia cepacia in patients with cystic fibrosis
Authors:David E Lacy  Anthony W Smith  Peter A Lambert  Daniel Peckham  David E Stableforth  EGrace Smith  Maya Desai  Peter H Weller  Michael RW Brown
Institution:Intensive Care Unit, Royal Liverpool Childrens Hospital, Alder Hey, Eaton Rd, Liverpool L12 2AP, UK;Department of Pharmaceutical Sciences, Aston University, Birmingham, UK;City Hospital, Nottingham, UK;Birmingham Heartlands Hospital, Birmingham, UK;Birmingham Children's Hospital, Birmingham, UK
Abstract:Early and accurate diagnosis of Burkholderia cepacia infection is important, particularly if segregation is to prevent patient-to-patient transmission. We have examined the serum response to a B. cepacia-specific 80-kDa outer membrane protein. 21 patients colonised with B. cepacia and Pseudomonas aeruginosa for 2–51 months (mean 11 months) were age- and sex-matched with 21 patients colonised with P. aeruginosa but not B. cepacia. The 80-kDa protein was recovered by electroelution from outer membrane proteins, separated by SDS-PAGE, coated onto ELISA plates, reacted with patient sera diluted 1:200, protein A-peroxidase and chromogenic substrate. We found that 19/24 patients colonised with B. cepacia and P. aeruginosa had high values, 2/24 patients had intermediate values, and 2/24 patients had a low value. 20/21 patients colonised with P. aeruginosa alone had low values and 1/21 had an intermediate value. We found that in the longitudinal serum samples studied from four patients only one patient developed high values after the first isolation of B. cepacia suggesting that seroconversion does not occur immediately after the first sputum culture of B. cepacia. We conclude that an ELISA test using B. cepacia-specific 80-kDa outer membrane protein can distinguish B. cepacia colonised and non-colonised patients and may be useful in the early diagnosis of B. cepacia infection.
Keywords:Outer membrane protein  Porin  Electroelution              Burkholderia cepacia
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