Detection of Legionella DNA by PCR of whole-blood samples in a mouse model.
A detection system for Legionella DNA in blood samples based on the PCR was developed and evaluated in A/J mice with experimentally induced Legionella pneumonia. Primers were designed to amplify a 106 bp DNA fragment of the 16S rRNA gene specific to Legionella species. The PCR system could detect clinically relevant Legionella species including Legionella pneumophila, Legionella micdadei, Legionella bozemanae, Legionella dumoffii, Legionella longbeachae, Legionella gormanii and Legionella jordanis. The sensitivity of the PCR system was 20 fg extracted DNA. In the mouse model, the blood PCR was compared with results obtained by PCR on bronchoalveolar lavage fluid (BALF) samples, cultures of blood and BALF and detection of Legionella urinary antigen. Blood PCR was positive until 8 days after infection, while BALF PCR became negative on day 4. These results indicate that PCR using blood samples may be a useful, convenient and non-invasive method for the diagnosis of Legionella pneumonia.
Aoki
S
S
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Hirakata
Y
Y
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Miyazaki
Y
Y
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Izumikawa
K
K
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Yanagihara
K
K
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Tomono
K
K
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Yamada
Y
Y
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Tashiro
T
T
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Kohno
S
S
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
Kamihira
S
S
Department of Laboratory Medicine1 and Second Department of Internal Medicine2, Nagasaki University School of Medicine, Nagasaki 852-8501, Japan.
D000818:Animals; D000942:Antigens, Bacterial / Q000652:urine; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis / Q000097:blood*; D004275:DNA, Ribosomal / Q000032:analysis / Q000097:blood; D004195:Disease Models, Animal; D007124:Immunoenzyme Techniques; D007875:Legionella / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007877:Legionnaires' Disease / Q000175:diagnosis* / Q000382:microbiology; D008297:Male; D051379:Mice; D008805:Mice, Inbred A; D016133:Polymerase Chain Reaction / Q000592:standards*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S