We report a case of cat scratch disease caused by Bartonella henselae in Korea. A 25-yr-old woman developed left cervical lymphadenopathy with history of contact with a dog. The cervical lymphadenopathy persisted for 1 month and resolved gradually and spontaneously. Serologic test was not done during the acute stage of the disease. Immunofluorescent antibody test performed during the convalescent stage was positive for B. henselae. To confirm B. henselae infection, polymerase chain reaction (PCR) analysis using aspirates of cervical lymph node was performed and the presence of B. henselae DNA was demonstrated. This is the first reported case of cat scratch disease in Korea confirmed by PCR for B. henselae DNA.
Adult ; Bartonella henselae/*genetics/*isolation and purification ; Cat-Scratch Disease/*diagnosis/*microbiology ; DNA, Bacterial/*analysis/*genetics ; Female ; Humans ; Polymerase Chain Reaction/*methods

Anti-Bacterial Agents ; therapeutic use ; Antibodies, Bacterial ; blood ; Bartonella henselae ; isolation & purification ; Cat-Scratch Disease ; diagnosis ; drug therapy ; microbiology ; Diagnosis, Differential ; Gentamicins ; therapeutic use ; Humans ; Lymph Nodes ; pathology ; Zoonoses

OBJECTIVETo establish polymerase chain reaction (PCR) technique for the detection of specific genes related to species of genus Bartonella, and for diagnosing clinically suspected cat-scratch disease (CSD) case complicated with pneumonia on both lungs. The appearance of Bartonella infectious diseases calls for genus and species detection and tools for identification in order to make clinical diagnosis and carry on epidemiological studies.

METHODSOne pair of primer TIle.455p-TAla.885n was designed based on the fact that tRNA(Ile)-tRNA(Ala) intergenic spacer region in 16S-23S rRNA intergenic spacer (ITS) of genus Bartonella were high variable sequences flanked by completely conserved tRNA-encoding genes. 16S-23S rRNA was longer than that which had been described in other bacteria. Two published pairs of primers were used to directly detect the specific gene fragments of Bartonella species DNA extracts from human blood, followed by PCR product Sequencing and nucleotide base sequence analysis.

RESULTSAmplification products of the three pairs of primers had the same predicted size of those in Bartonella spp. According to the different length of electrophoresis bank, the sample was identified as a species of genus Bartonella other than the positive control. Sequence analysis showed that the nuleotide sequence from the PCR product of primer TIle.455p-TAla.885n was identical to the Bartonella isolated from Yunnan in China.

CONCLUSIONSPCR-based assay provided a simple and rapid means to detect pathogenic Bartonella species in humans and mammalian hosts as well as in arthropod vecters. This study suggested that this pathogenic Bartonella species existed in patients in northern and southern parts of China.

Animals ; Bartonella ; genetics ; isolation & purification ; Bartonella Infections ; diagnosis ; microbiology ; Base Sequence ; Cat-Scratch Disease ; diagnosis ; microbiology ; Cats ; China ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; RNA, Transfer, Ala ; chemistry ; genetics ; RNA, Transfer, Ile ; chemistry ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult