Detection of Neisseria meningitidis group A, Haemophilus influenzae type b, and Streptococcus pneumoniae antigens in cerebrospinal fluid specimens by antigen capture enzyme-linked immunosorbent assays

Abstract

Antigen capture enzyme-linked immunosorbent assay was compared to coagglutination and counterimmunoelectrophoresis for the detection of meningococcal, Haemophilus, and pneumococcal antigens. Enzyme-linked immunosorbent assay detected 1 ng of purified meningococcal and Haemophilus polysaccharides per ml and 5 ng of pneumococcal polysaccharide per ml; coagglutination detected 20, 25, and 30 ng/ml, respectively, of these polysaccharides; and counterimmunoelectrophoresis detected 10, 50, and 60 ng/ml. Double-antibody sandwichantiglobulin enzyme-linked immunosorbent assays, which employed antibodies produced in two animal species, differentiated 100% of the cerebrospinal fluid (CSF) specimens from meningococcal meningitis patients and 95% of the CSFs from Haemophilus patients from heterologous control CSFs. Double-antibody sandwich procedures, which use the same antiserum preparation for coating the wells of microtiter plates and for alkaline phosphatase-conjugated immunoglobulin, differentiated meningococcal CSFs from control specimens but were unable to effectively differentiate the Haemophilus or pneumococcal specimens from control CSFs. Coagglutination detected specific antigen in 92% of the meningococcal CSFs, 80% of the Haemophilus CSFs, and 92% of the pneumococcal specimens. The comparable percentages for counterimmunoelectrophoresis were 76, 95, and 71%.

Laboratory diagnosis of bacterial meningitis.

“…The addition of a substrate for the enzyme results in the production of a colored product if specific bacterial antigen was present in the CSF specimen and bound to the primary antibody. EIAs have been evaluated for their abilities to detect H. influenzae type b, S. pneumoniae, and N. meningitidis antigens in CSF (12,37,134,142,162). The sensitivities and specificities of these tests have been reported to be 84 to 100% and 89 to 100%, respectively.…”

Comparison of a new, rapid enzyme-linked immunosorbent assay with latex particle agglutination for the detection of Haemophilus influenzae type b infections

“…Previous investigators have noted the potential sensitivity and specificity of ELISA systems for the detection of haemophilus PRP (2,4,7,9,13). In addition, ELISA has theoretical advantages over LPA, including nonsubjective endpoints.…”

“…Emphasis has been placed on the sensitivity of these tests, since failure to treat haemophilus infection promptly could have disastrous consequences. Thus, tests such as counterimmunoelectrophoresis, which cannot detect the low concentrations of PRP present in some invasive H. influenzae type b infections (1,5,(9)(10)(11), have largely been abandoned in favor of more sensitive tests, such as latex particle agglutination (LPA). LPA can detect 1 ng or less of PRP per ml in clinical specimens and has been found to identify nearly all culture-documented haemophilus infections (3,(8)(9)(10)(11)(12).…”

Pertussis: the disease and new diagnostic methods.

“…These include counterimmunoelectrophoresis (CIE), latex agglutination, and coagglutination (11). Sensitive ELISAs have been developed for detection of Haemophilus influenzae (11,37,145), Streptococcus pneumoniae (62,145), and Neisseria meningitidis (145) antigens in clinical specimens, using polyclonal or monoclonal antibodies. Various enzyme immunoassays have also been developed for detection of viral and bacterial antigens in respiratory secretions (2,27,101,171), cerebrospinal fluid (152), and endocervical and urethral specimens (122,152).…”