尖吻蝮蛇舌形虫若虫粗抗原诊断效果及组分分析  

Diagnostic effects and fraction of crude antigens from Armillifer agkistrodontis nymphs

作　　者：张玲玲[1] 陈家旭[1] 陈韶红[1] 蔡玉春[1] 郭俭[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾,血吸虫病和丝虫病合作中心,上海200025

出　　处：《中国人兽共患病学报》2012年第4期367-370,379,共5页Chinese Journal of Zoonoses

基　　金：国家科技支撑计划(2008BAI56B03);国家科技重大专项(2008ZX10004-011;2009ZX10004-302;2012ZX10004220)

摘　　要：目的分析尖吻蝮蛇舌形虫若虫粗抗原的蛋白组成、免疫特性及在舌形虫病诊断中的诊断效果。方法自感染尖吻蝮蛇舌形虫虫卵4个月的小鼠体内取尖吻蝮蛇舌形虫若虫,匀浆,用盐析法制备抗原。以此粗制抗原包被微量反应板,用间接ELISA方法检测尖吻蝮蛇舌形虫病人、感染小鼠及其它寄生虫病人血清中的特异性IgG抗体。用聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹(Western Blot)技术分析该粗制抗原的有效抗原蛋白组分。结果 ELISA方法检测尖吻蝮蛇舌形虫病人血清5份,正常人血清17份,感染小鼠、血吸虫病、肺吸虫病、肝吸虫病、囊虫病、旋毛虫病病人血清各20份,蛔虫病、钩虫病、鞭虫病病人血清各10份,发现舌形虫病人和感染小鼠血清均为阳性,其它寄生虫病人血清均为阴性。经SDS-PAGE和Western Blot分析,尖吻蝮蛇舌形虫成虫粗抗原可见5条主带和14条次带(Mr16-200kDa);若虫粗抗原可见9条主带和13条次带(Mr16-150kDa),其中有16条特异性条带可被感染小鼠血清识别,5条特异性条带可被舌形虫病人血清识别,与肺吸虫病、鞭虫病病人血清在约Mr34kDa附近处出现较微弱的交叉反应带,同其它寄生虫病人血清未出现交叉反应带。结论尖吻蝮蛇舌形虫若虫粗抗原同其它寄生虫病的交叉反应较小,可用于尖吻蝮蛇舌形虫病的血清学检测,而其有效的诊断抗原成分需经纯化后再行进一步的分析。The purpose of the present study was to analyze the component and immunological properties of the crude antigens from A.agkistrodontis nymphs and observe the diagnostic effects of crude antigens in pentastomiasis.The A.agkistrodontis nymphs were collected from the mice on 120 days post-infection and washed with saline.Then,they were manually homogenized by using a glass homogenizer,and the crude antigens were extracted by centrifugation.The crude antigens,as a coating antigen,were used to detect the specific IgG antibodies in sera from patients of pentastomiasis and other parasitic diseases,and from mice infected with A.agkistrodontis eggs by indirect ELISA.The effectively antigenic components of crude antigens were analyzed by SDS-PAGE and Western Blot.The indirect ELISA results revealed that the sera of patients of pentastomiasis and mice infected with A.agkistrodontis eggs were all positive,whereas the sera of patients with other parasitic diseases（20 paragonimiasis,20 shistosomiasis,20 clonorchiasis,20 cysticercosis,20 trichinellosis,17 normal,10 ascariasis,10 hookworm disease,10 trichuriasis,respectively） were all negative.The crude antigens of adults were separated into 19 distinct bands（Mr16-200 kDa） in SDS-PAGE gel;the crude antigens of nymphs were separated into 22 distinct bands（Mr16-150kDa） and displayed 16 distinct bands and 5 immune reaction bands recognized by mixed serum of mice and patients of pentastomiasis respectively on Western Blot.Besides,it was at about Mr 34kDa where found much slightly cross-reactions with paragonimiasis and trichuriasis,and there was no cross-reaction with others.In conclusion,the crude antigens of A.agkistrodontis nymphs have little cross-reaction with other parasitic diseases and are valuable for serological detection of pentastomiasis.However,the effectively antigenic components need to be purified and further analysis should be conducted.

关 键 词：舌形虫病 尖吻蝮蛇舌形虫 粗抗原 ELISA WESTERN BLOT 

分 类 号：R384.9[医药卫生—医学寄生虫学]