血平板-纸桥法在沙门氏菌血清分型中的应用探究  

Study on the application of blood plate-paper bridge method in Salmonella serotyping

作　　者：涂晓波 马淑棉 金晓蕾 匡燕云 刘慧玲 牛娜 赵芳 吕敬章 林燕奎 TU Xiao-bo;MA Shu-mian;JIN Xiao-lei;KUANG Yan-yun;LIU Hui-ling;NIU Na;ZHAO Fang;LV Jing-zhang;LIN Yan-kui(Food Inspection and Quarantine Technology Center of Shenzhen Customs District,Shenzhen,Guangdong 518000,China;不详)

机构地区：[1]深圳海关食品检验检疫技术中心,广东深圳518000 [2]深圳市检验检疫科学研究院

出　　处：《现代预防医学》2024年第22期4184-4190,共7页Modern Preventive Medicine

基　　金：食品安全国家标准立项计划(spaq-2019-50)。

摘　　要：目的综合分析血平板-纸桥法进行H抗原诱导和鉴定的影响因素,探究其机理,以促进该方法在沙门氏菌血清分型中的应用。方法通过比较不同纸桥材质对H抗原诱导和鉴定的影响,采用扫描电镜对不同纸桥材质表面进行扫描,探究其影响机理,并结合环境湿度和生理盐水用量等因素,优化沙门氏菌血清分型的血平板-纸桥法实验条件。采用优化后的方法与GB 4789.4标准的简易平板法,对40株沙门氏菌进行血清型鉴定进行比较。结果CN110和CN95硝酸纤维素膜的哑光面与普通滤纸的过桥效果相近,差异无统计学意义(P>0.05),而过滤滤膜效果显著低于普通滤纸(P<0.05)。沙门氏菌在更高湿度下蔓延效果更好,80%湿度下与50%、20%组的结果相比存在显著差异(P<0.0167)。随着生理盐水量的增加,蔓延效果显著提升(P<0.001),最优量是15μL。使用两种方法对40株食源性沙门氏菌进行鉴定,本研究方法能一次性准确鉴定沙门氏菌血清型,而简易平板法需1至3次诱导。结论本研究所优化的血平板-纸桥法操作简便且诱导效果显著,适合于沙门氏菌血清型的鉴定。此方法可作为沙门氏菌血清分型方法的有效补充,提高食源性疾病的溯源效率和准确性。Objective To comprehensively analyze the influencing factors of the blood plate-paper bridge method for H antigen induction and identification,and to investigate its mechanism in order to promote the application of this method in Salmonella serotyping.Methods The experimental conditions of the blood plate-paper bridge method for Salmonella serotyping were optimized by comparing the effects of different paper bridge materials on the induction and identification of H antigen,scanning the surface of different paper bridge materials by scanning electron microscope to explore the mechanism of the effects,and combining the factors of environmental humidity and saline dosage.The optimized method was compared with the simple plate method of GB 4789.4 standard for serotyping 40 strains of Salmonella.Results The matte surface of CN110 and CN95 nitrocellulose membranes had similar bridging effect with ordinary filter paper,and the difference was not statistically significant(P>0.05),while the effect of filter membrane was significantly lower than that of ordinary filter paper(P<0.05).Salmonella spread better at higher humidity,and there was a significant difference between the results at 80%humidity compared with those of the 50%and 20%groups(P<0.0167).The spreading effect was significantly improved with increasing amount of saline(P<0.001),and the optimal amount was 15μL.Forty foodborne Salmonella strains were identified using the two methods,and the method in this study was able to accurately identify Salmonella serotypes in one go,whereas the simple plate method required one to three inductions.Conclusion The blood plate-paper bridge method optimized in this study is easy to operate and has a significant induction effect,which is suitable for the identification of Salmonella serotypes.This method can be used as an effective complement to Salmonella serotyping methods to improve the efficiency and accuracy of foodborne disease traceability.

关 键 词：血平板-纸桥法 沙门氏菌 血清分型 

分 类 号：R-33[医药卫生]