Detection of humoral response using a recombinant heat shock protein 70, DnaK, of Mycoplasma haemofelis in experimentally and naturally hemoplasma-infected cats.

Hemoplasmas is a trivial name given to a group of erythrocyte-parasitizing bacteria of the genus Mycoplasma. From hemoplasmas cat, Mycoplasma haemofelis is the Fungal Recombinant Proteins most pathogenic, while "Candidatus Mycoplasma haemominutum" and "Candidatus Mycoplasma turicensis" less pathogenic. shotgun library of genomic DNA is fragmented M. haemofelis built, and the colonies randomly selected for sequencing DNA. In silico-translated amino acid sequence of the open reading frame than the alleged mass spectrometry data of M.

 haemofelis spots identified as immunogenic proteins by two-dimensional gel electrophoresis and Western blotting. Three of the spots matched with predictable sequence of heat shock protein 70 (DnaK) homolog, elongation factor Ts, and a fragment of phosphoglycerate kinase that is found during screening library. 

A full-length copy of the M. haemofelis DnaK gene was cloned into Escherichia coli and recombinant. M. haemofelis purified recombinant DnaK and then used in Western blotting and enzyme-linked immunosorbent assay (ELISA) to investigate the humoral immune response during acute infection in cats experimentally infected with M. haemofelis, "Mycoplasma Ca. haemominutum," or "Mycoplasma turicensis Ca. ". 

Recombinant M. haemofelis DnaK ELISA was Conjugates Recombinant Proteins  also used to screen clinical samples submitted for testing hemoplasma PCR to a commercial laboratory (n = 254). Experiments infected cats become seropositive following infection, with greater and earlier antibody response seen in cats inoculated with M. haemofelis than those seen in cats inoculated with "Mycoplasma Ca. haemominutum" or "Ca. Mycoplasma turicensis," both by Western blotting and ELISA. From a clinical sample, 31.1% had antibodies detected by ELISA but only 9.8% were positive by PCR for one or more hemoplasmas.