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 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 and biofilm formation related genes of Staphylococcus aureus in pigeon slaughter and processing. The enterotoxin contents and biofilmforming abilities of 41 strains of Staphylococcus aureus were determined using the Staphylococcus aureus enterotoxin enzyme-linked immunoassay kit and crystal violet staining method. Meanwhile, 12 enterotoxin genes and 10 biofilm formation-related genes of Staphylococcus aureus were also amplified by PCR. Among the 41 strains of Staphylococcus aureus, 30 strains were enterotoxin-producing strains in enterotoxin enzyme-linked immunoassays. In the detection of enterotoxin genes, 35 strains of bacteria contained enterotoxin genes (with the detection rate being 85.36%), and have carried one or more enterotoxin genes. The detection rates of classical enterotoxin genes weresea (29.27%), seb (29.27%), sec (4.88%), sed (65.85%) and see (2.44%), respectively; The detection rates of novel enterotoxin genes were sel (36.59%), seg (29.27%), sem (17.07%), seu (17.07%), sei (14.63%), she (7.32%) and sek (0.00%), respectively. The detection rates of the strains with weak, moderate and strong biofilm-forming abilities determined by crystalline violet staining were 39.02%, 43.90% and 17.07%, respectively. In the detection of biofilm formation related genes, the detection rates of clfB, luxS and clfA were the highest (all at 100.00%), 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, and bap was not detected. In addition, all strains were found to carry multiple biofilm-related genes, and the number of the strains carrying 7 genes was the highest (15/41, 36.59%). The Staphylococcus aureus contaminated in the pigeon slaughtering and processing had more enterotoxins and higher biofilm-forming ability. It was predicted that the Staphylococcus aureus found in the pigeon slaughter and processing was pathogenic and had potential risk of causing food poisoning.