多重PCR对金黄色葡萄球菌杀白细胞素基因的检测

目的应用多重PcR检测含Panton-Valentine杀白细胞素（PVL）基因的金黄色葡萄球菌。方法收集我院2005年1月至2006年1月从临床多种标本中分离的金黄色葡萄球菌，用多重PCR同时检测葡萄球菌16SrRNA基因、mecA基因和lukS/F-PV基因。多重PCR检测MRSA的SCCmec基因型及亚型。结果195株金黄色葡萄球菌经多重PCR检测，121株为耐甲氧西林金黄色葡萄球菌（MRSA），74株为甲氧西林敏感金黄色葡萄球菌（MSSA），共检测到26株金黄色葡萄球菌lukS/F-PV基因阳性，阳性率为13．3％（26，195）。其中19株为MRSA，阳性率为15．7％（19／121）；7株为MSSA，阳性率为12．2％（7／74）。19株lukS/F-PV基因阳性的MRSA的SCCmec基因型分别为SCCmec Ⅲ型10株、SCCmecⅢA型4株、SCCmecⅣ型4株及SCCmecⅠ型1株。26株lukS/F-PV基因阳性的分离株有11株分离自脓液或创面分泌物，10株分离自痰标本，3株分离自血液标本，2株分离自尿液。结论在温州地区分离的MRSA和MSSA中都能检测到Panton-Valentine杀白细胞素基因，含Panton-Valentine杀白细胞素基因MRSA的SCCmec基因型主要为SCCmec Ⅲ，含PVL基因的金黄色葡萄球菌主要引起化脓性感染和肺部感染。

Multiplex PCR for rapid detection of Panton-Valentine leukocidin gene from clinical isolates of Staphylococcus aureus and its clinical significance

Objective To detect Panton-Valentine leukocidin gene (PVL gene) from the isolates of Staphylococcus aureus. Methods One hundred and ninety-five strains of Staphylococcus aureus were isolated. PVL gene of the isolates was determined by multiplex PCR. The genotypes of SCCmec were also determined by another multiplex PCR. Results Among 195 isolates, 121 isolates were methicillin-resistant (MRSA) and 74 were methicillin-sensitive Stapkylococcus aureus (MSSA). Twenty-six of 195(13.3%) isolates were lukS/F-PV genes positive. Nineteen of 121(15.7%) isolates of MRSA and 7 of 74(12.2%) were lukS/F-PV positive. The genotypes and subgenotypes of staphylococcal chromosomal cassette mec of 19 isolates of MRSA carrying lukS/F-PV genes was SCC mec III (10 isolates), SCCmec III A(4 isolates), SCCmec IV (4 iaolates) and SCCmec I(1 isolates) . Twenty-six isolates carrying lukS/F-PV genes were isolated from abscess (11 isolates), sputum (10 isolates) , blood (3 isolates) and urine (2 isolates). Conclusion Genes coding for PVL are encountered in MRSA and MSSA isolates. SCCmec III are major genotypes in MSRA carrying lukS/F-PV gene. Staphylococcus aureus carrying PVL gene are associated with pyogenic infection and pneumonia.