Detection of Staphylococci in Mouse Phagocytic Cells by in Situ Hybridization Using Biotinylated Dna Probes
作者:
MatsuhisaAkio,
SaitoYoshihiro,
UeyamaHiroshi,
AikawaYouko,
OhonoTsuneya,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1994)
卷期:
Volume 69,
issue 1
页码: 31-37
ISSN:1052-0295
年代: 1994
DOI:10.3109/10520299409106258
出版商: Taylor&Francis
关键词: intracellularStaphylococci;phagocytes;nonradioisotopic detection;in situ hybridization;avidin-biotin technology;neutrophils;septic diagnosis;bacteriolytic enzymes;cell morphology
数据来源: Taylor
摘要:
In situ hybridization was used to detect intracellularStaphylococcus aureusandS.epidermidisin mouse phagocytic cells after experimental infection of C3H mice withStaphylococcivia abdominal or intravenous injection. Isolated ascites or whole blood were tested by the phagocyte smear technique, using bacteriolytic enzymes to preserve phagocytic cell morphology. The exposed bacterial DNA was visualized as intracellular hybridized signals by use of biotinylated DNA probes and by immunocytochemistry using streptavidin-alkaline phosphatase conjugates as detector molecules. These DNA probes, prepared from randomly cloned genomic DNA fragments ofS. aureusand S.epidermidis, were strain-specific and did not cross-hybridize either in situ or on dot-blot hybridization. This technique of in situ hybridization with phagocyte smears is useful for detection and diagnosis of intracellular bacteria regardless of viability.
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