ABSTRACT
Objective: To identify the monoclonal antibody specific to Aeromonas spp., a Gram negative bacteria causing gastroenteritis and wound infection. Methods: The monoclone, namely 88F2-3F4, was produced from hybridoma technology. The specificity of antibody secreted from 88F2-3F4 was tested against other Gram negative bacteria frequently found in gastrointestinal tract. Then the antibody was used for searching Aeromonas antigens in artificial seeded rectal swab cultures by dot-blot enzyme linked immunosorbent assay. Results: 88F2-3F4 produced an antibody that recognized an antigen with a molecular mass of 8.5 kDa in all 123 isolates of the seven Aeromonas species tested, but recognized no epitope of any other Gram-negative bacterium typically found in the gastrointestinal tract. A dot-blot enzyme linked immunosorbent assay based on this antibody showed 86.49% sensitivity and 92.13% specificity. Conclusions: 88F2-3F4 monoclonal antibody could react with all Aeromonas isolates, but not other Gram negative bacteria, therefore it should be a useful tool for the detection of Aeromonas antigen in clinical and environmental samples.
ABSTRACT
This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.