摘要
目的分析河北省一起鼠疫疫情的实验室判定流程,探讨河北省鼠疫疫情实验室判定的标准化操作程序。方法对2017年10月康保牧场捡获的自毙鼠解剖取材,按腺、肝、脾、肺、心顺序,分别进行细菌培养,同时做噬菌体裂解实验;再取肝、脾剪碎制成悬液,应用胶体金免疫层析实验检测鼠疫菌F1抗原,压印载玻片2张(一张革兰氏染色镜检,一张美兰染色镜检),同时留3份肝、脾材料(分别用于抗原检测、基因检测和备用),以备后续实验。结果捡获的5份自毙鼠材料有3份胶体金实验和反相血凝试验都出现阳性,效价高达1∶10 240以上,PCR检测Fra和Pla基因,抗原阳性的3份标本都扩增出目标条带,细菌培养3份均阳性。结论准确判定了此次鼠疫疫情,同时成功建立了一套适应《鼠疫诊断标准(ws279-2008)》的实验室诊断技术流程。
Objective To study the laboratory determination process of a plague epidemic event in Hebei Province,and try to establish a standard operation procedure for lab determination of plague epidemic situation.Methods Self-dead rats collected from Kangbao Pasture in Oct.2017 were dissected,and the organs,such as glands,liver,spleen,lung,and heart were used for bacteria culture and phage lysis experiments.Liver and spleen were cut into pieces and made into suspension used for the following tests immediately:Colloidal gold immunochromatographic assay(CGIA) for detecting plague bacillus antigen F1;Print two glass slides(one for gram staining check,and the other one for methylene blue staining check);Triplicate specimens of liver and spleen were prepared for latter use(antigen test,gene test and reserve).Results Among the specimens of 5 self-dead rats,3 showed positive results both in CGIA,and the following reverse phase hemagglutination test,and the titers were as high as 1∶10 240 and above.In PCR tests for genes Fra and Pla of the 3 specimens,target bands were achieved.In bacteria culture,results of the 3 specimens were positive.Conclusion The event was determined as a plague epidemic,and a lab diagnostic technical process meeting plague diagnostic standards(ws279-2008) has been establisheation.
作者
杨晓燕
杜国义
刘合智
闫东
陈永明
周松
王海峰
YANG Xiao-yan;DU Guo-yi;LIU He-zhi;YAN Dong;CHEN Yong-ming;ZHOU Song;WANG Hai-feng(Hebe Provincial Plague Institute, Zhangjiakou 075000, China)
出处
《中华卫生杀虫药械》
CAS
2019年第2期141-143,共3页
Chinese Journal of Hygienic Insecticides and Equipments
基金
2017年度河北省医学科学研究重点课题计划(编号:20170468)
国家重点研发计划课题(编号:2016YFC1201304)
关键词
鼠疫菌
自毙鼠
检测技术
鼠疫判定
plague bacillus
self-dead rat
test technology
plague determination
