鸽屠宰加工环节金黄色葡萄球菌肠毒素、生物膜形成能力及其相关基因的检测

Detection of Staphylococcus aureus enterotoxins, biofilm-forming ability and their associated genes in pigeon slaughtering and processing

摘要:

为了解鸽屠宰加工环节中金黄色葡萄球菌的肠毒素、生物被膜形成能力、肠毒素及生物被膜形成相关基因的情况。采用金黄色葡萄球菌肠毒素酶联免疫分析试剂盒和结晶紫染色法检测41株金黄色葡萄球菌的肠毒素含量和生物被膜形成能力，同时通过PCR方法扩增金黄色葡萄球菌12种肠毒素基因和10种生物被膜形成相关基因。41株金黄色葡萄球菌在肠毒素酶联免疫法检测中，有30株为产肠毒素的菌株。肠毒素基因的检测中，有35株菌含肠毒素基因，检出率为85.36%，且有携带一种或多种肠毒素基因的情况。经典肠毒素基因的检出率分别为：sea（29.27%）、seb（29.27%）、sec（4.88%）、sed（65.85%）和see（2.44%）；新型肠毒素基因的检出率分别为：sel（36.59%）、seg（29.27%）、sem（17.07%）、seu（17.07%）、sei（14.63%）、seh（7.32%）和sek（0.00%）。结晶紫染色法测定菌株生物被膜形成能力弱、中等和强的检出率分别为39.02% 、43.90%和17.07%。生物被膜形成相关基因的检测中，clfB、luxS和clfA的检出率最高均为100.00%，其次是sarA和ccp均为97.56%，sigB、cna、icaA、agrC的检出率分别为85.37%、65.85%、41.46%、2.44%，bap未检出。此外，所有菌株均有携带多种生物被膜相关基因的情况，以携带7种基因的菌株最多（15/41，36.59%）。鸽屠宰加工环节污染中金黄色葡萄球菌存在较多的肠毒素且生物被膜形成能力较高，预测该鸽屠宰加工环节中金黄色葡萄球菌具有致病性，存在引起食物中毒的潜在风险。

Abstract:

To understand the enterotoxins, biofilm forming ability, enterotoxins and biofilm formation related genes of Staphylococcus aureus in pigeon slaughter and processing. Determination of enterotoxin content and biofilm formation ability of 41 strains of Staphylococcus aureus using Staphylococcus aureus enterotoxin enzyme-linked immunoassay kit and crystal violet staining method,12 enterotoxin genes and 10 biofilm formation-related genes of Staphylococcus aureus were also amplified by PCR. 41 strains of Staphylococcus aureus with 30 enterotoxin-producing strains in enterotoxin enzyme-linked immunoassays. In the detection of enterotoxin genes, 35 strains of bacteria contained enterotoxin genes, the detection rate was 85.36%, and have carried one or more enterotoxin genes. The detection rates of classical enterotoxin genes were, respectively: sea（29.27%）、seb（29.27%）、sec（4.88%）、sed（65.85%）and see（2.44%）; The detection rates of novel enterotoxin genes were, respectively: sel（36.59%）、seg（29.27%）、sem（17.07%）、seu（17.07%）、sei（14.63%）、seh（7.32%）and sek（0.00%）. The detection rates of weak, moderate and strong biofilm forming ability of the strain as determined by crystalline violet staining were 39.02%, 43.90% and 17.07%, respectively. Biofilm formation-related genes in the assay, clfB, luxS and clfA had the highest detection rate of 100.00% each, followed by sarA and ccp both at 97.56%, the detection rates of sigB, cna, icaA, and agrC were 85.37%, 65.85%, 41.46%, and 2.44%, respectively, bap not detected. In addition, all strains were found to carry multiple biofilm-related genes, the highest number of strains carrying 7 genes (15/41, 36.59%). Major of enterotoxins and higher biofilm formation capacity of Staphylococcus aureus in pigeon slaughter and processing contamination, predicted pathogenicity of Staphylococcus aureus in the slaughter and processing of this pigeon, potential risk of food poisoning.