Detection of Meticillin-resistant Staphylococcus aureus by Fluorescence in situ Hybridization and Flow Cytometry
- VernacularTitle:荧光原位杂交-流式细胞术法检测耐甲氧西林金黄色葡萄球菌
- Author:
Yijun ZHU
;
Xiangyang LI
- Publication Type:Journal Article
- Keywords:
Meticillin-resistant Staphylococcus;
Fluorescence in situ hybridization;
Flow cytometry;
Detection method;
PCR
- From:
Chinese Journal of Nosocomiology
1994;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To establish and evaluate detection method of meticillin-resistant Staphylococcus aureus(MRSA) by fluorescence in situ hybridization(FISH) and flow cytometry.METHODS The experiment conditions including bacteria suspension concentrations,probe concentrations,and hybridization temperatures were optimized for FISH test in the light of orthogonal design principle.The fluorescence signals were detected by flow cytometer,and detection results by FISH and flow cytometry were compared with that of real-time PCR.RESULTS The optimal hybridization conditions of FISH detecting MRSA were 40?105 of bacteria suspension concentrations per microliter of hybridization buffer solution,and 4 ng of probe concentrations per microliter of hybridization buffer solution and 50 ℃ of hybridization temperatures.The sensitivity and specificity of detecting MRSA by FISH-FCM were 97.3% and 100.0%,respectively,and detection results between FISHFCM and real-time PCR had not significant deviation.CONCLUSIONS Fluorescence in situ hybridization and flow cytometry,which detect directly mecA gene in MRSA, are a fast and accurate methods for MRSA detection and can apply in clinical laboratory.
