酵素抗体法による緑膿菌菌体抗原検出に関する基礎的並びに臨床的検討

Abstract

A sandwich enzyme-linked immunosorbent assay (ELISA) for detection of a 11 serotypes of P. aeruginosa has been developed by using polyclonal rabbit anti P. aeruginosa (NC-5 strain) antiserum. The following results were obtained: 1. Cultured broth of P. aeruginosa was detected most sensitively by this assay followed by sonicated soluble antigen, washed bacterial fluid and heated soluble antigen of P. aeruginosa. The lower limit of detection was approximately 2.3 X 10(4) cfu/ml of P. aeruginosa (NC-5 strain) (containing 22.2 ng/ml of protein) in cultured broth. 2. By this assay, all serotypes of P. aeruginosa and some strains of Family Pseudomonadaceae (RNA group I) were detected, but no cross reaction was noticed to other species of bacteria (24 species, 268 strains). 3. The serum antigen in the experimental sepsis of P. aeruginosa or murine experimental pneumonia was detected when at least approximately 10(3) cfu/ml of P. aeruginosa was present in serum or BALF. But the antigen was detected from concentrated urine even under the concentration of 10(2) cfu/ml in sepsis. 4. P. aeruginosa antigens in sputum of patients with chronic respiratory tract infection was also detected. There was a significant difference between absorbance value of sputum in non-infected patients (group 1) and one in infected patients by P. aeruginosa (group 3) (p less than 0.01). And there was a strong correlation between absorbance value of sputum in Group 3 and number of P. aeruginosa. In 22 of 33 sputa (66.7%) with P. aeruginosa infection as verified by conventional culture methods, the antigen was detected in sputum. In a patient of P. aeruginosa pneumonia with acute leukemia, the antigen was detectable from sputum and one of the series of concentrated urine. It is concluded that the detection of P. aeruginosa antigen in sputum and urine by ELISA is a helpful aid to diagnose P. aeruginosa infection.