An ELISA method for detecting unexpected antibodies.

R M Aloisi; J C Dowd

Home/Archive/PubMed

PubMed · 7032295

An ELISA method for detecting unexpected antibodies.

R M Aloisi·J C Dowd

View original source ↗Export metadata ↓

Abstract

This paper reports a new horseradish peroxidase enzyme linked immunosorbant assay (ELISA) to detect patient serum alloantibodies. The new methodology utilizes a water-soluble chromogen, O-dianisidine and a temporary solid antibody support, osmotically lysed reagent erythrocytes. In a double blind study the horseradish peroxidase ELISA was found to be as sensitive as the conventional antiglobulin test. In parallel dilution studies on serum samples containing anti-Kell antibodies, the horseradish peroxidase-ELISA was found to be substantially more sensitive than the conventional antiglobulin test. While the procedure is at present impractical for routine use, these results indicate a potential for improved sensitivity which warrants further research. Possible applications and prospective areas of research are discussed.

Explore related subjects

Antibodies, Anti-Idiotypic Coombs Test Dianisidine Double-Blind Method Enzyme-Linked Immunosorbent Assay Humans Immunoenzyme Techniques Immunoglobulin G Isoantibodies Kell Blood-Group System Comparative Study Journal Article

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

R M Aloisi, J C Dowd. 1981. An ELISA method for detecting unexpected antibodies.. https://pubmed.ncbi.nlm.nih.gov/7032295/

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

Application of a combined protocol for rational request and utilization of antibody assays improves clinical diagnostic efficacy in autoimmune rheumatic disease.

CONTEXT: Because of a marked increase in the number of requests for antinuclear antibodies, anti-extractable nuclear antigen antibodies, and anti-double-stranded DNA antibodies for the diagnosis of autoimmune rheumatic disease, guidelines have been proposed for their appropriate use. OBJECTIVE: To evaluate in terms of clinical efficacy and cost-benefit ratio the outcome of applying a protocol for the diagnosis of autoimmune rheumatic disease. DESIGN: A diagnostic protocol for the rational utilization of second-level tests (anti-extractable nuclear antigen antibodies and anti-double-stranded DNA antibodies) was applied at Hospital Polyclinic beginning January 2004. The appropriateness of 685 consecutive requests received at the clinical pathology laboratory from January to June 2004 was assessed. Patients who underwent these laboratory tests were followed up for 12 months after blood sample drawing. RESULTS: Introduction of the protocol led to a significant reduction in the number of second-level tests prescribed (27.9% vs 49.5% for anti-extractable nuclear antigen antibodies; 27.5% vs 56.6% for anti-double-stranded DNA antibodies). After the period of observation, none of the 163 patients who had negative results on the first-level test and were asymptomatic, for whom second-level tests had not therefore been performed, were found to have autoimmune rheumatic disease. In 90.5% (77/85) of patients positive for the second-level tests, clinical confirmation of autoimmune rheumatic disease was obtained. CONCLUSIONS: Not only did application of the diagnostic protocol reduce the number of second-level tests performed but it also increased their specificity. Our data thus indicate that the use of shared guidelines by clinical and laboratory specialists yields satisfactory results.

Antibodies, Anti-Idiotypic↗

PubMed2006

Humoral immune response and immunoglobulin G Fc receptor genotype are associated with better clinical outcome following idiotype vaccination in follicular lymphoma patients regardless of their response to induction chemotherapy.

We have reported that anti-idiotype antibody response and FcgammaRIIIa 158 valine/valine (V/V) genotype both correlate with better outcome in a group of 136 follicular lymphoma patients receiving idiotype vaccination after induction chemotherapy. Here, we examined whether this correlation is related in any way to the chemotherapy response. In patients with complete response (CR), the 5-year progression-free survival (PFS) was 69% for patients with antibody response and/or V/V genotype, while the PFS was only 40% for patients with neither; the median time to progression (TTP) was 10.47 versus 3.46 years (P=.012). In patients with partial response (PR), the 5-year PFS was 57% for patients with antibody response and/or V/V genotype, and 17% for patients with neither; the median TTP was not reached versus 1.31 years (P=.001). This study further confirms the strong association of clinical outcome with antibody response and with the functionally more active form of the Fc receptor in patients receiving idiotype vaccination regardless of their response to induction chemotherapy.

Antibodies, Anti-Idiotypic↗

PubMed2006

Human beta-defensin-3 induction in H. pylori-infected gastric mucosal tissues.

AIM: To examine human beta-defensin-3 (hBD-3) expression in inflamed gastric mucosal tissues or MKN45 gastric cancer cells with or without H. pylori infection for better understanding the innate immune response to H. pylori. METHODS: We used reverse transcription-polymerase chain reactions and immunohistochemistry to examine hBD-3 expression in inflamed gastric mucosal tissues or MKN45 gastric cancer cells with or without H. pylori. Effects of hBD-3 against H. pylori were also evaluated. RESULTS: The mean mRNA expression of hBD-3 in H. pylori-positive specimens was significantly higher than that in H pylori-negative specimens (P = 0.0002, Mann-Whitney). In addition, unlike uninfected samples, 8 of 15 (53.33%) infected mucosal samples expressed hBD-3 protein. H. pylori dose-dependently induced mRNA expression of hBD-3 in MKN45 cells, an effect inhibited by adding anti-toll-like receptor (TLR)-4 antibody. HBD-3 protein completely inhibited H. pylori growth. CONCLUSION: Our results suggest that like hBD-2, hBD-3 may be involved in the pathophysiology of H. pylori-induced gastritis.

Antibodies, Anti-Idiotypic↗