基于微球液态阵列分子技术的沙门菌血清分型研究

Study on Salmonella serotyping by use of Microsphere-based Liquid Array method

目的 了解基于微球液态阵列分子技术对广东省腹泻病例沙门菌分离株的分型效果。方法 在微球液态阵列分子平台上应用SSA试剂盒对沙门菌人源株进行分子血清分型。结果 2010-2014年广东省人源沙门菌4 942株分为189种血清型，前100种血清型占所有菌株的98.08%（4 847/4 942），其中98%可用SSA试剂盒完整分型；采用SSA试剂盒检测198株菌O抗原，可检出181株；用SSA检测H抗原的结果（98.32%，528/537）与传统血清凝集试验结果相符；fljB基因的符合率为93.09%（175/188），fljB基因假阴性率为7.35%（9/134），假阳性率为7.41%（4/54）；sdf基因和Vi基因符合率均为100%；用SSA试剂盒检测12株血清不能分型的沙门菌，有11株能成功分型。结论 SSA试剂盒能对96%以上的广东省人源沙门菌株进行分子血清分型，其结果与传统方法符合率超过98%。基于微球液态阵列分子技术的血清分型法比传统方法更具有高通量且快速的特点。

Objective To understand the effect of serotyping on Salmonella isolates, by use of Microsphere-based Liquid Array method, among diarrhea patients, in Guangdong. Methods Salmonella isolated from humans in Guangdong province were serotyped on the Microsphere-based Liquid Array platform with SSA kit. Results A total of 4 942 Salmonella strains with 189 serotypes, were identified in Guangdong province in 2010-2014. The top 100 serotypes accounted for 98.08% （4 847/4 942） of all the strains. 98% of the top 100 species serotypes could completely be serotyped with SSA kit. In order to detect O antigen among 198 isolates with SSA kit, 181 strains were carrying the O antigen, with the coincidence rate as 100%. However, under the SSA, 98.32% （528/537） of the H antigen could be detected and were consistent with the traditional serum agglutination test. The coincidence rate of fljB gene was 93.09% （175/188）, with false negative rate and false positive rate of fljB gene as 7.35% （9/134） and 7.41% （4/54） respectively. The coincidence rate of sdf gene and Vi gene were 100%. 11 out of the 12 Salmonella strains could not be serotyped under the traditional methods but were successfully serotyped by the molecular serotyping method. Conclusions Using the SSA kit, more than 96% of the anthropogenic Salmonella strains could be serotyped in Guangdong province. Comparing with the traditional methods, the coincidence rate of serotyping appeared over 98%. Under the Microsphere-based Liquid Array techniques, the molecular serotyping method appeared faster and more accurate on Salmonella serotyping than those traditional methods.