Molecular Detection of Infectious Endocarditis (Bartonella quintana) Bacteria from Selected Military Hospitals

 Bartonella quintana is an aerobic, gram-negative, rod-shaped, and polar bacterium. Detection of this bacterium is done through blood culture in an agar medium, and the longtime of detection by culture has made molecular methods such as PCR important for more accurate and faster detection.

 For this reason, 100 cultured negative endocarditis specimens were collected in this study. DNA extraction was performed from B. quintana, and the concentration and quality of the obtained DNA were measured. PCR reaction was performed on the genome of negative control samples. To clone a portion of the amplified gene in PUC 18 plasmid, the PCR product was first purified. After ligation, JM107 E. coli susceptible to calcium chloride was used. Transformed bacteria were cultured on LB Broth medium containing Ampicillin antibiotic. Then 2 to 3 white colonies were selected, and PCR was performed. Plasmid extraction was performed after confirming the presence of recombinant and inserted plasmids.

The last dilution of PUC18 plasmid for B. quintana with an initial concentration of 780 ng/µL, which formed a detectable band on the gel, was calculated to be 10-7, and the minimum number of detectable copies in a 25 μL PCR reaction equal to 24 copies. . In quantitative DNA analysis, its amount was calculated between 1.69 and 1.8.

 The collected samples were then examined for the presence of B. quintana in patients. Of the 60 samples collected, none were positive.