甘肃省鼠疫自然疫源地鼠疫耶尔森菌耐链霉素基因rpsL突变位点的检测  

Detection of rpsL mutations in streptomycin resistant gene of Yersinia pestis from different natural foci in Gansu Province

作　　者：郭丽民[1] 袁育 何爱伟 吴斌[1] 苗克军[1] 王鼎盛[1] 徐大琴[1] 席进孝[1] 王扬扬 安文静 GUO Limin;YUAN Yu;HE Aiwei;WU Bin;MIAO Kejun;WANG Dingsheng;XU Daqin;XI Jinxiao;WANG Yangyang;AN Wenjing(Gansu Provincial Center for Disease Control and Prevention,Lanzhou,Gansu 730020,China;Luqu center for Disease Control and Prevention,Luqu,Gansu 747200,China)

机构地区：[1]甘肃省疾病预防控制中心,甘肃兰州730020 [2]碌曲县疾病预防控制中心,甘肃碌曲747200

出　　处：《中华卫生杀虫药械》2024年第6期559-563,共5页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：甘肃省自然科学基金(22JR11RA185)。

摘　　要：目的分析甘肃省鼠疫疫源地鼠疫耶尔森菌(鼠疫菌)耐链霉素基因rpsL序列突变特征,为突发人间鼠疫病临床用药及鼠疫菌耐药监测提供依据。方法选取1962—2022年分离自甘肃省不同类型鼠疫疫源地376株鼠疫菌,提取DNA,根据鼠疫菌耐链霉素基因rpsL设计PCR引物和TaqMan-MGB荧光探针,采用PCR产物直接测序法分析rpsL基因的突变位点,实时荧光PCR技术对甘肃省不同疫源地鼠疫菌耐链霉素基因rpsL的128位点碱基突变进行检测。结果376条rpsL基因序列比对分析发现甘肃省不同类型鼠疫疫源地鼠疫菌rpsL基因全序列均未发生变异,不同年代的鼠疫菌rpsL基因碱基序列均相同,证实甘肃省鼠疫菌rpsL基因序列保守。376株甘肃省不同鼠疫疫源地鼠疫菌羧基荧光素(FAM)探针检测结果均为阳性,对应检测rpsL基因(128:A),ΔRn(normalized reporter)>3000000;所有鼠疫菌株亚磷酰胺(VIC)探针检测结果均为阴性,对应检测rpsL基因(128:G),ΔRn<100000。结论建议甘肃省喜马拉雅旱獭疫源地动物鼠疫持续流行的鼠疫监测点实验室进行鼠疫菌耐药性监测,及时发现本地鼠疫菌rpsL基因特征,为突发人间鼠疫诊疗提供依据。Objective To detect the mutations of rpsL gene in strains of Yersinia pestis from different natural foci in Gansu Province,so as to provide basis for clinical medication of the plague patients and monitoring on drug resistance of Yersinia pestis.Methods DNA was extracted from 376 strains of Yersinia pestis isolated from different natural foci of Gansu province from 1962 to 2022.PCR primers and TaqMan-MGB fluorescent probes were designed according to the rpsL mutations of Yersinia pesti s with the streptomycin resistant.The mutation sites of the rpsL gene were analyzed by direct sequencing of PCR products,and the 128 base mutations in the rpsL gene of Yersinia pestis from different foci of Gansu Province were detected by Real-time PCR.Results It was found that the entire rpsL gene sequence of Yersinia pestis from different types of plague foci in Gansu Province did not change through the sequence comparison analysis of 376 rpsL gene,and the rpsL gene base sequence of Yersinia pestis from different years was the same,which confirmed that the rpsL gene sequence of Yersinia pestis in Gansu Province was conserved.FAM test results of 376 strains of Yersinia pestis from different natural foci in Gansu Province were positive,corresponding to rpsL(128:A),ΔRn>3000000;The VIC test results of all strains were negative,corresponding to rpsL(128:G),ΔRn<100000.Conclusion It is recommended to monitor the drug resistance of Yersinia pestis in the Himalayan Marmota foci of Gansu Province where the animal plage continues to spread,so as to find out the mutations of rpsL gene in Yersinia pestis in time,and provide basis for the diagnosis and treatment of human plague.

关 键 词：鼠疫菌 RPSL基因 TaqMan-MGB 检测 

分 类 号：R378[医药卫生—病原生物学]