PubMed · 11020312
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T T Ngo. 2000. Developments in immunoassay technology.. https://doi.org/10.1006/meth.2000.1030
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We have evaluated the prevalence of HIV-1 non-B subtypes in Spain by means of an enzyme immunoassay (EIA) for discrimination between B and non-B subtypes. Samples were obtained from newly diagnosed patients attended at internal medicine outpatient clinics between October 1997 and October 1998. Discrimination between HIV-1 B and non-B subtypes was carried out by means of the EIA, with V3 synthetic peptides specific to the different subtypes. Non-B-serotyped samples were genetically analyzed in the gp41 region from the original sera. During the study period, 909 samples were collected from 21 medical units located in various Spanish geographical regions. Serotyping was possible in 885 cases, of which 791 were assigned as B serotype (89.38%), 70 showed no reactivity to any of the peptides (7.91%), and the remaining samples displayed other reaction patterns (2.72%). Of the 94 non-B-assigned samples, 65 were genetically characterized in the gp41 region of the env gene: 55 were B subtype, 5 were A subtype (4 clustered with CRF02AG reference strains), 3 were C subtype, and 2 were G subtype. The prevalence rate for non-B subtypes in Spain was established at 1.13% (95% CI, 0.59-2.21). Although the B subtype is predominant in the Spanish population, other subtypes have been detected.
The HLA-B27 antigen is an important genetic marker in ankylosing spondylitis (AS). Methods for the detection of B27 include the microlymphocytotoxicity test and, more recently, flowcytometry (FC). Here, we describe a new method, IMS-ELISA, for measuring the B27-antigen. It combines immunomagnetic separation (IMS), to obtain B27-positive cells from whole blood samples, with an enzyme-linked immunosorbent assay (ELISA) as a read-out. IMS-ELISA was tested on 367 samples obtained from five different hospitals in Taiwan. The sensitivity, specificity and accuracy of the method were compared with FC. Any conflicting data between IMS-ELISA and FC was confirmed by HLA-DNA typing via PCR-SSP (polymerase chain reaction-sequence specific primers). Overall, the results for sensitivity, specificity and accuracy obtained by IMS-ELISA and FC did not show any significant difference (p>0.05). However, when considering laboratory time, cost, ease of operation and the screening of large samples for HLA-B27, the IMS-ELISA was superior to the FC method. We conclude that IMS-ELISA may be used as a fast screening method for HLA B27 detection.
OX40 (CD134) is a member of the TNF receptor family and is expressed selectively on activated T lymphocytes. Through interactions with OX40 ligand, OX40 delivers potent costimulatory signals to T cells. Here, we describe a sandwich ELISA method for the detection and quantification of OX40 using the monoclonal anti-OX40 antibodies ACT35 and L106 as capture and detecting antibodies, respectively. With this ELISA, the existence of naturally occurring soluble forms of OX40 (sOX40) could be demonstrated for the first time. Soluble OX40 is detectable in serum of subpopulations of healthy donors and patients with autoimmune disease and cancer. Among the five diseases that were investigated, chronic lymphocytic leukemia was identified as the one with the highest frequency of sOX40-positive sera and with the highest mean sOX40 serum concentration. The stability of sOX40 is not affected by repeated freeze/thaw cycles nor by prolonged storage at -20 degrees C, allowing studies on the potential diagnostic significance of sOX40 using existing banks of sera.