Serological diagnosis of experimental Enterococcus faecalis endocarditis.
A modified rat model of endocarditis with catheterization for 2 days was established in female Lewis rats using different inocula of Enterococcus faecalis (strain no. EF 19) in order to measure IgG antibodies in serum during the course of infection. Increasing the inocula intravenously resulted in an increase in the CFU/g vegetation and the CFU/g spleen, the ID50 being about 10 CFU/ml and the ID90 about 1x10(2) CFU/ml. The lowest bacterial inoculum infecting 100% of the rats was 3x10(3) CFU/ml, and for further investigations we used this inoculum size. Rats were sacrificed on day 2, 5, 7, 9, 11 and 28 after infection. The CFU/g vegetation and the CFU/g spleen increased until day 7 and then decreased. Serum samples were collected from 129 rats at different times after challenge. Three different ELISA systems were established to measure the IgG antibody responses: E. faecalis sonicate ELISA (a pool of four sonicates of strain no. EF 10, EF 11, EF 19 and EF 48), E. faecalis whole cell ELISA (strain no. EF 19) and E. faecalis purified cell wall ELISA (strain no. EF 19). An IgG antibody response was detected already on day 2, and except for a minor decrease on day 6/7 the antibody response continued to increase until day 14 (whole cell ELISA and sonicate ELISA) and day 21 (purified cell wall ELISA) when a plateau was reached. Significant increases in IgG antibody responses (p<0.05) were found between groups of rats from days 0-2, 2-8/9 and 8/9-14 in the E. faecalis whole cell and sonicate ELISAs and from days 0-2, 2-10/11 and 10/11-21 in the E. faecalis purified cell wall ELISA. In conclusion, we established a model of endocarditis in rats with catheterization for 2 days and were able to demonstrate an increase in IgG antibodies during the course of infection.
Kjerulf
A
A
Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.
Espersen
F
F
Gutschik
E
E
Majcherczyk
P A
PA
Hougen
H P
HP
Rygaard
J
J
Høiby
N
N
D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D002473:Cell Wall / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D004697:Endocarditis, Bacterial / Q000175:diagnosis* / Q000276:immunology / Q000473:pathology; D013293:Enterococcus faecalis / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis* / Q000276:immunology / Q000473:pathology; D006351:Heart Valves / Q000382:microbiology / Q000473:pathology; D051381:Rats; D011917:Rats, Inbred Lew