Urdaneta H, Guimarães S, Silva E F, Tavares C A
Department of Biology, Los Andes University, Venezuela.
Rev Inst Med Trop Sao Paulo. 1994 Nov-Dec;36(6):539-45. doi: 10.1590/s0036-46651994000600011.
A sensitive and specific Capture Sandwich ELISA (CSE) was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG) against E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.
利用针对三种不同无菌培养的溶组织内阿米巴菌株（来自巴西的CSP和来自墨西哥的HM1-IMSS）的多克隆纯化兔抗体，开发了一种灵敏且特异的捕获夹心酶联免疫吸附测定法（CSE），用于检测粪便样本中的粪抗原。通过两个阶段从用滋养体全提取物免疫的兔子中分离出抗溶组织内阿米巴的免疫球蛋白G（IgG）：首先在含有与琼脂糖4B结合的溶组织内阿米巴抗原的柱上进行亲和层析，随后在琼脂糖抗体4B-蛋白A上进行另一次层析。使用纯化抗体的捕获夹心酶联免疫吸附测定法能够检测到70纳克的阿米巴蛋白，灵敏度为93%，特异性为94%。显微镜检查和CSE的联合检测一致性和不一致性分别为93.25%和6.75%。得出的结论是，CSE对于检测感染患者粪便中溶组织内阿米巴的粪抗原具有高度特异性，执行速度更快，比显微镜检查更容易且更灵敏。
