旋毛虫病的免疫诊断研究

旋毛虫病的免疫诊断研究刘光明蔡海松综述宋思扬苏文金审校厦门大学生物学系（厦门361005）旋毛虫病的临床表现错综复杂，临床诊断较为困难，易与其它传染病相混淆。肌肉活检发现幼虫或囊包虽可确诊，但在轻度感染和感染早期往往不易检出，即使是感染晚期，因受摘取..

XPS Dissection of the Optical Transition Additive in Farm Film

报道了无保护气氛条件下,采用高温固相还原法合成适用于农用塑料地膜的光转换添加剂,共掺铜、铕激活剂的硫化钙(CaS)无机荧光材料,用X射线光电子能谱(XPS)方法对光转换添加剂中不同工艺下激活剂元素进行表征,再通过光致发光谱、透射光谱测量,探索材料中激活剂的能量传输特性,为高新农业的发展提供清洁能源。An optical transition additive used in farmfilmwas synthesized by doping the Cu and Eu activators into an inorganic fluorescent material,CaS with the high-temperature solid phase reduction method under unˉprotected atmosphere.The characteristics of the activators in the films prepared under different conditions were measured by X-ray photo-electron spectroscopy.The energy-transfer properties of the activators in the films were investigated by the photo-induced luminescence spectra and transmission spectra.It is useful for developing a clean energy source in agriculture.福建省自然科学基金资助项目(A0110006);; 厦门大学自选课题基金资助项目(Y07007

PURIFICATION of P49 ANTIGEN GENE PRODUCT of TRICHINELLA SPIRALLS EXPRESSED IN ESCHERICHIA COLI

目的建立旋毛虫P49抗原基因原校表达产物的纯化方法。方法菌体经冻融、超声破菌及提取包涵体后，用离子交换和凝胶过滤层析纯化蛋白质。结果纯化后的融合蛋白P49/gST经SdS-PAgE电冰鉴定达电泳纯。双抗体夹心ElISA法检测结果表明纯化的P49/gST融合蛋白能与旋毛虫感染的鼠血清和纯化融合蛋白免疫的兔血清反应，而不与正常民和兔血清反应。结论纯化后的融合蛋白P49/gST具有较高的纯度及较强的免疫活性，可望作为旅毛虫病的免疫诊断抗原。Aim To establish a purification procedure for p49 antigen gene product of Trichinella spiralis expressed inEschedchia coli.Methods After centrifugation of the bacterial culture, the bacterial pellet was resuspended and lysed byfreezing-thawing treatment and ultrasonication.Ion exchanged and gel filtration chromatography were used to purify the fu-sion protein p49/GST from the inclusion bodies- Re8ultS The purity of the fusion protein p49/GST was verified using SDS-PAGE.The purified p49/GST protein could react with mouse antisera against Tricinella sPiralis and rabbit antisera againstpurified p49/GST protein, but not with normal mouse and rabbit sera by sandwich EL1SA.Concluslon The purified fusionprotein p49/GST with high purity and good immune activity could be used for the immune aasay of trichinellosis.福建省重点（农医）项目!95－Z－15

DIAGNOSIS OF TRICHINOSIS BY ELISA WITH P49/GST ANTIGEN

目的以纯化的融合蛋白 p49/GST为抗原建立ELISA检测方法。方法对一批试验血清进行间接ELISA检测。结果 19份人工感染鼠血清、5份人工感染猪血清、4份旋毛虫病猪血清、4份病人血清呈IgG抗体阳性 ,2 1份人工感染鼠血清呈IgM抗体阳性 ,而正常对照血清及 30 0份屠宰场待检猪血清均呈阴性反应 ,其结果与常规压片法结果相符。结论融合蛋白p49/GST对于研制旋毛虫病的诊断抗原具有的潜在应用价值Aim To establish ELISA detecting method for the specific antibodies of Trichinella spiralis.Method A series of confirmed trichinosis sera were detected with ELISA with fusion protein p49/GST.Results Positive results of IgG antibody were found in 19sera of experimental infected mouse and 3sera of infected swine and 4 sera of patients;positive results of IgM antibody were found in 21 sera of experimental infected mouse .All normal sera were negative. Conclusion The ELISA with p49/GST can be regarded as a sensitive, specific immunological method for the diagnosis of trichinosis.福建省重点(农医 )项目资助!(项目号 :95 -Z -15 0

A TWO-PARTICLE TURBIDIMETRIC LATEX IMMUNOASSAY FOR THE DETECTION OF SPECIFIC ANTIBODIES OF TRICHINELLA SPIRALIS

目的　建立旋毛虫病的快速检测技术。方法　基因工程抗原包被有色乳胶颗粒，抗抗体包被磁性颗粒，在抗体存在下形成抗原－ 乳胶－ 抗体－ 抗抗体－ 磁性颗粒的复合场，在磁场作用下沉淀下来，从而达到快速检测旋毛虫相关抗体的目的。结果　１９ 份人工感染鼠血清、５ 份人工感染猪血清、３ 份旋毛虫病猪血清、４ 份病人血清均呈阳性反应，而对照血清均为阴性反应；该方法在鼠感染旋毛虫后第五天可测出ＩｇＭ 抗体，第九天可测出ＩｇＧ抗体。结论　本检测技术简便易行，不需专用设备，可望成为一种快速诊断旋毛虫病的有效方法。Aim To establish a rapidly detecting method for the specific antibodies of Trichinella spiralis Method Latex was coated with p49/GST and polystyrene beads(Dynabeads)were coated with anti-antibodies SAM-IgG、SAH-IgG、RAP-IgG or SAM-IgM Then the test serum was incubated with two particles for 30 min at room temperature with slowly shaking After sedimentation of the polystyrene beads with a magnet,the turbidity of the resultant latex suspension was estimated with eyes or measured spectrophotometrically at a wavelength of 400nm Results Positive result of IgG antibody were found in 19 sera of experimental infected mice,3 sera of experimental infected swine and 4 sera of patients The IgM and IgG can be check up in mice after 5 and 9 days after experimental infection The result agreed with those obtained by ELISA Conclusion A two-particle turbidimetric latex immunoassy established by this study is a rapid,easy and precise method for the diagnosis of the trichinosis福建省科委(农医)项目!(95-Z150

The Cloning of complementary DNA encoding p49 ES antigen of Trichinella spiralis

应用dnA重组技术将编码旋毛虫肌幼虫49kdA抗原的基因克隆于大肠杆菌中.设计特异的PCr引物，用PT-PCr技术直接从旋毛虫肌幼虫总rnA中反转录并扩增出约1.1kb的靶dnA.用bAMHI和ECOrI酶解后将其克隆到质粒载体PuC19中，用X-gAl培养基筛选重组子.该重组子经相同的核酸内切酶水解获得约2.7kb和1.1kb两个片段，分别与PuC19和目的基因的大小相同，目的基因经序列分析并与文献报道的序列比较发现具有97.8%的同源性.Complementary DNA(cDNA) encoding 49KD ES antigen of Trichinella spiralis muscle larvae was cloned in E.coli by recombinant DNA technology.Aabout1.1kb DNA fragment was obtained by RT PCR from total RNA of Trichinella spiralis using a specific 5′ end 3′ end primer.PCR product was cleaved with restriction enzymes EcoRI and BamHI and ligated into the pUC19 vector.The cloning was screened by X gal medium.When the recombinant DNA was cleaved by the same enzynes, 1.1kb and 2.7kb fragments were obtained, which were similar to the langth of PCR product and of pUC19 vcvtor.There is 97.8 percent homologous between the sequence of PCR product and the sequence which reported in literature by sequencing analysis.福建省重点（农医）资