间接血凝试验在囊虫病诊断中的应用

囊虫病是人畜共患疾病,严重损害人民健康。本文报道用间接血凝试验检测人血清中囊虫抗体,具有较高的特异性和敏感性。 1 囊虫抗原致敏血球的制备 1·1 囊虫抗原的制备,选取新鲜的含猪囊蚴的猪肉,用消毒空针抽取囊液,经3000rpm/分离心10min,上清即为囊液抗原。用分光光度计测量蛋白     

抗囊虫卵黄免疫球蛋白的提取

猪囊虫病是严重危害人体健康和养殖业发展的重要人兽共患病。医学和兽医科技工作者对检测猪囊尾蚴病进行了大量的研究 ,已取得了一定的进展。但在以卵黄IgY作为抗体方面尚未有人尝试。本试验旨在探索一个提取抗囊虫抗体的新方法。为囊虫病的防治及检测开辟一条新的途径。1　猪囊虫抗原的纯化1 1　材料　猪囊虫含生理盐水 15 0ml。1 2　方法1 2 1　囊液的收集　从新鲜囊虫猪肉摘取虫囊 ,然后将虫体取出置灭菌平皿中 ,用生理盐水洗涤 4次 ,吸干多余液体。用剪刀剪破囊壁使囊液自然流出 ,收集囊液于灭菌瓶内 ,置4℃过夜。次日将囊液做 30 0 0…     

包虫与囊虫间共同抗原的实验探讨

本文用E.gCF和C.cCF致敏血球检测了四种抗原(E.gCF、E.gPS、C.cCF和C.cSCW)免疫的小白鼠。结果发现:囊虫头节及囊壁不仅是囊虫特异性抗原的载体而且是与包虫发生交叉反应的共同抗原的主要来源;原头节共同抗原较少。包虫囊液特异性抗原性比囊虫囊液要强。     

包虫和猪囊虫抗原免疫化学性质及酶联免疫印斑技术临床应用研究

用 SDS 聚丙烯酰胺凝胶电泳和酶联免疫印斑技术分别对人源细粒棘球蚴囊液、棘球蚴砂、猪囊尾蚴囊液抗原组分进行分析研究,并对两种病血清学交叉反应进行了试验。结果显示包虫囊液抗原有29条蛋白区带,有两条糖蛋白带。猪囊虫囊液抗原显示36条蛋白带谱,两条糖蛋白带。棘球蚴砂显示23条蛋白带谱。包虫病人血清与印有包虫囊液抗原的硝酸纤维膜反应显示23条反应带,其中特异带为6条。包虫病人、猪囊虫病人血清与此膜反应显示4条共同反应带。包虫病人血清与棘球蚴砂抗原膜反应显示4条主要特异反应带,包虫病人血清、猪囊虫病人血清与此膜反应显示1条共同反应带。猪囊虫病人血清与猪囊虫囊液抗原膜反应显示22条反应带,其中有5条特异反应带;包虫病人和囊虫病人血清与此膜反应显示3条共同反应带。     

ELISA检测脑囊虫病人脑脊液循环抗原

应用酶联免疫吸附试验(ELISA)检测脑囊虫病人循环抗原(CAg)一些学者已做了不少工作。我们于1990年用 ELISA 检测瞄囊虫病人脑脊液 CAg 获得了较好的效果。一、材料与方法(一)抗原制备自新鲜病猪肉摘取囊尾蚴,抽取囊液,经离心和上清液透析后为抗原,保存于-20℃备用。(二)兔抗囊尾蚴抗体制备取囊尾蚴囊液抗原多点注射健康兔,效价在 ELISA 1∶5000时,取血分离血清,分装冰冻干燥,置于-20℃备用。(三)脑脊液(CSF) 脑囊虫病患者的 CSF 均采自本所根据临床症状、皮下结节活检及头颅 CT 扫描等确诊的住院病人,共68份。其他脑部疾病(病毒性脑炎、脊     

ELISA法间接检测囊虫患者血清特异抗原的探讨

目的 探讨简便的检测囊虫抗原的方法。方法 囊液抗原包被酶标板，ELISA法检测待检血清与含ELISA法1：8( )的囊虫抗体兔血清的混合液的抗体，滴度低于1：2(#)为阳性。结果 囊虫患者阳性率在82．69％-85．37％之间。讨论囊液抗原间接检测囊虫患者血清特异抗原结果稳定，方法简便，临床符合率较高。     

高压液相色谱法分离提纯囊虫抗原的研究

<正> 囊虫病的免疫及免疫诊断的重要问题之一是抗原问题,本文首次应用Beckman344M型高压液相色谱仪分离制备囊虫不同部位抗原物质,应用ELISA试验,检测所分离的各不同组分的抗原活性,其结果报告如下。 材料和方法 一、制备粗抗原 按常规收集囊液制备囊液粗抗原(简称CF_0Ag)再分别制出全囊虫粗抗原(CW_0Ag)和头节粗抗原(CS_0Ag),分别测其蛋白含量后-20℃保存。 二、高压液相色谱法分离制备抗原     

阿苯达唑对猪囊尾蚴糖原作用的观察

1980年我国首先应用阿苯达唑临床治疗囊虫病,大量病例临床疗效观察证实,该药对皮肌型和脑型囊虫病均有显著疗效。本文报道阿苯达唑对虫体糖原的作用,借以探讨该药对囊尾蚴的作用机理。 材料与方法:猪囊尾蚴取自武汉市肉类联合加工厂当天宰杀的新鲜“米猪肉”,选择大小正常,充满透明囊液的虫体,在生理盐水中漂洗2次,置40％生理     

免疫亲和层析法提取猪囊尾蚴抗原的鉴定及其在血清学诊断中的应用价值

本文采用免疫亲和层析法,提取与自然感染猪囊虫的猪血清IgG反应的猪囊虫抗原。所提取头节抗原、囊壁抗原和囊液抗原的Desc-PAGE区带分别为7、13和10条,其中囊液PAS染色显示3条阳性带(层析前为4条);SDS-PAGE区带分别为12、15和13条。相对分子量为65、53、40和30～29KDa的抗原含量最多,免疫原性最强。提取的抗原应用于免疫电泳,可提高其敏感性。     

猪囊尾蚴液、固相多种抗原对囊虫病免疫诊断效果评价

目的　筛选适合多种寄生虫病流行区使用的特异性较高的囊虫病免疫诊断抗原。　方法　制备猪囊尾蚴各类型液相抗原和固相抗原 ,用ELISA或IEST检测囊虫病流行区患者阳性血清抗体 ,比较各种抗原的敏感性和特异性。　结果　猪囊尾蚴全囊及囊液尿素溶性抗原与水溶性纯化抗原ELISA检测囊虫抗体的特异性差异无显著性 (P >0 .0 5 )。固相抗原IEST有敏感性好、操作简易的特点。　结论　猪囊尾蚴全囊尿素溶性纯化抗原是较理想的囊虫病ELISA诊断抗原 ,有诊断应用价值。固相石腊切片抗原IEST较为适合流行区现场或基层应用。     

SPA-ELISA评价三种不同纯度牛源细粒棘球蚴囊液抗原在包虫病免疫诊断中的价值

本文采用SPA-ELISA法评价了三种不同纯度牛源细粒棘球蚴囊液抗原在包虫病免疫诊断中的价值,用三种抗原检测了41例包虫病患者血清中特异IgG抗体的水平,它们的阳性率分别为:牛源细粒棘球蚴液粗抗原86.0％,半饱和硫酸铵法纯化抗原91.4％,氯化镁—磷钨酸法纯化抗原92.3％,两种纯化抗原与粗抗原的阳性检出率相比,经统计学处理具有显著差异,认为西北地区牦牛体内寄生的细粒棘球蚴囊液抗原不乏抗原活性;氯化镁—磷钨酸法和半饱和硫酸铵法是两种简便易行,纯化效果理想的抗原纯化方法,宜于在基层推广应用。     

脑囊虫病人血清中循环抗原特性的分析

本研究用免疫印渍法，以三种抗血清为探针，对脑囊虫病人血清中的循环抗原、囊虫头节和体壁抗原及囊液抗原进行了比较观察。结果表明循环抗原与两种囊虫抗原相同的组分有４个ＫＤ数为７５、４３、４０、２３的多肽；仅与虫头及囊壁相同的抗原为６５ＫＤ多肽；仅与囊液相同的多肽为８５、１９ＫＤ二组分。这显示循环抗原主要来源于虫囊的全部结构；循环抗原可诱发三种特异性免疫球蛋白的产生，依次为ＩｇＧ、ＩｇＡ及ＩｇＭ。可诱发ＩｇＧ的循环抗原多肽是８５、５３、４３、４０、３１、１９、１８．５及１７．５ＫＤ多肽；可诱发ＩｇＡ抗体的抗原多肽ＫＤ数为６５、４９、４３、４０及３５的组分；而ＩｇＭ特异性抗体产生于ＫＤ数为７５、４３、４０及３１的多肽。三种抗体血清用于检测血清中的循环抗原都显示出良好的应用价值。免疫兔血清的敏感性与特异性均不亚于单克隆抗体的识别结果。４３及４０ＫＤ多肽是循环抗原中的主要组分，是诊断的重要指标。     

SPA-ELISA检测卡氏肺孢子虫抗原的研究

目的 探讨SPAK-ELISA检测卡氏肺孢子虫抗原对诊断卡氏肺孢子虫肺炎的价值。 方法 采用SPA-ELISA检测免疫抑制大鼠模型在不同时期肺泡灌洗液(BALF)和血清中肺孢子虫(PC)抗原,以及造血干细胞移植并发肺炎患者痰液和血清抗原,并与病原学检查结果比较。 结果 感染大鼠于用药后第4周,BALF中PC抗原,以及肺组织印片和BALF涂片包囊检查开始出现阳性,但前者阳性率(60％)高于后者(20％和40％),而且全部阳性达到的时间(8周)也早于后者(10周)。第10周有2只鼠血清抗原阳性,但此后又转阴。30例肺炎患者痰液抗原阳性8例,阳性率为26.67％,8例抗原阳性患者中只有3例发现包囊,符合率为37.5％。血清抗原检测全部阴性。 结论 SPA-ELISA检测痰液和肺泡灌洗液抗原比病原学检查敏感性高,而检测血清抗原不适用于诊断卡氏肺孢子虫肺炎。     

Inhib-ELISA与SPA-ELISA检测人及不同种属动物血清抗体的比较研究

本文以鼠疫病人、鼠疫感染狗及旱獭血清作为实验模型,比较观察了Inhib-ELISA与SPA-ELISA检测不同种属血清抗体的敏感性。结果表明,在检测不同种属血清抗体的敏感性和特异性方面,Inhib-ELISA优于SPA-ELISA。Inhib-ELISA检测血清抗体不受种属特异性影响,而SPA-ELISA仍存在一定的局限性。因此,Inhib-ELISA更适宜于人兽共患病血清流行病学检测工作。     

Diagnosis of internal acariasis with avidin-biotin system enzyme-linked immunosorbent assay

AIM: To explore the value of avidin-biotin system enzymelinked immunosorbent assay (ABC-ELISA) in diagnosis of intestinal acariasis.METHODS: Mite-specific IgG levels in serum of 48 patients with intestinal acariasis were measured with ABC-ELISA.The sensitivity of this method was compared with that of staphylococcal protein A enzyme-linked immunosorbent assay (SPA-ELISA).RESULTS: The positive rate of mite-specific IgG detected with ABC-ELISA and SPA-ELISA was 89.58% (43/48) and 56.25% (27/48), respectively. The positive rate with ABCELISA was statistically higher than that with SPA-ELISA (X2=13.50, P＜0.01).CONCLUSION: ABC-ELISA is an effective method for the diagnosis of intestinal acariasis.     

Clinical usefulness of determining beta-2-microglobulin concentration in thyroid cystic fluid

The main subject of the paper was to prove that beta-2-microglobulin (beta-2-m) exists in fluid taken from a thyroid gland cyst of the patients with non-toxic, nodular goitre with cysts and whether the confirmation of beta-2-m existence in fluid taken from a cyst may indicate the presence of autoimmunological process in thyroid glands. The research was carried out on a group of 36 patients operated on because of goitre. Patients with the thyroid cancer were except. The operations were carried out in the General Surgery Department of Hospital 1 in Sosnowiec from 1994 to 1999. The group of patients consisted of women aged between 13 to 75 years old (average 43 +/- 13.8 years). TSH, FT4 serum concentration in this investigated group were normal. The paper presents the beta-2-m existence in fluid taken from a thyroid gland cyst. It was confirmed that there was no differences between beta-2-m serum concentration in operated patients and in healthy controls. In 50% of patients operated on with thyroid cysts high antibody antithyroid peroxidase (antiTPO) serum concentration and in fluid taken from cyst was confirmed and statistically higher beta-2-m concentration in cyst fluid than beta-2-m concentration in serum. CONCLUSIONS: The increased antiTPO concentration in serum and in a fluid taken from cyst and beta-2-m existence in cyst fluid point to the autoimmunological process taking place in half of the patients with thyroid cysts.     

Diagnosis of intestinal acariasis with avidin-biotin system enzyme-linked immunosorbent assay

AIM: To explore the value of avidin-biotin system enzyme-linked immunosorbent assay (ABC-ELISA) in diagnosis of intestinal acariasis. METHODS: Mite-specific IgG levels in serum of 48 patients with intestinal acariasis were measured with ABC-ELISA. The sensitivity of this method was compared with that of staphylococcal protein A enzyme-linked immunosorbent assay (SPA-ELISA). RESULTS: The positive rate of mite-specific IgG detected with ABC-ELISA and SPA-ELISA was 89.58% (43/48) and 56.25% (27/48), respectively. The positive rate with ABC-ELISA was statistically higher than that with SPA-ELISA (chi2=13.50, P<0.01). CONCLUSION: ABC-ELISA is an effective method for the diagnosis of intestinal acariasis.     

Accumulation of hormones in breast cyst fluid.

The concentration of some immunoreactive peptide hormones in breast cyst fluid and serum from a series of patients with cystic breast disease were compared. The concentrations of the two pituitary gonadotropins, LH and FSH, were consistently higher in serum than in cyst fluid, confirming earlier more limited observations. The mean PRL concentration, on the other hand, was elevated in cyst fluid though the variability was great. The cyst fluid serum ratio of hCG was elevated in the patient group as a whole. However, the results suggested the presence of two subpopulations; group I, in which the cyst fluid and serum hCG levels were essentially indistinguishable and in the range found in normal serum, and group II, in which the cyst fluid hCG concentration was consistently greater than the normal serum range. Analysis of hormone concentrations in multiple cysts aspirated at the same time showed a remarkable degree of agreement in the cyst hormone levels in most of the patients. The greatest variability in the levels was seen with hCG, were a small numer of subjects showed a very large spread in the values.     

Immunoheterogeneity of parathyroid hormone in parathyroid cysts: diagnostic implications

Parathyroid cysts are uncommon lesions of the neck leading to hypercalcemia in a significant percentage of cases. The distinction between parathyroid and thyroid cysts is difficult to make on a clinical basis alone and relies on the demonstration of elevated PTH levels in cyst fluid. We describe a case of a parathyroid cyst in which intact PTH (1-84) levels were misleadingly low while midmolecule 44-68 PTH was markedly elevated. To explain this discrepancy, we studied cyst fluid from this and two other patients using Sephadex G-75 gel chromatography. Fractions were analyzed using an immunoradiometric assay for intact hPTH (1-84) and a RIA specific for the midmolecular 44-68 region of hPTH. Immunoreactivity corresponding to hPTH (1-84) was absent in the first case but present in the remaining two. Immunoreactive peaks corresponding to PTH fragments were demonstrable in all three cyst samples. Patients with elevated hPTH (1-84) in cyst fluid were hypercalcemic; in contrast, the patient with a low cyst level of hPTH (1-84) was normocalcemic despite having markedly elevated levels of midmolecule PTH (44-68) in both serum and cyst fluid. Parathyroid cysts may thus produce fragments rather than intact PTH; reliance on an intact hPTH assay could lead to misdiagnosis. The measurement of PTH by a midmolecular assay may be preferable to the measurement of intact PTH in the evaluation of fluid from cystic neck masses.     

Functional similarities of hepatic cystic and biliary epithelium: studies of fluid constituents and in vivo secretion in response to secretin

Hepatic cysts are a frequent manifestation of autosomal dominant polycystic kidney disease, but little is known about their functional characteristics. The goals of our study were to define the composition of hepatic cyst fluid and to determine whether hepatic cysts secrete in response to intravenously administered secretin. We percutaneously punctured five hepatic cysts and one proximal renal cyst from six subjects with autosomal dominant polycystic kidney disease and one solitary hepatic cyst from a subject without autosomal dominant polycystic kidney disease. Most fluids had an electrolyte composition similar to serum. Fluid from all hepatic cysts had glutamyltranspeptidase concentrations above those found in serum [( cyst]/[serum] = 4.93 +/- 5.92), contained secretory component (the epithelial receptor for polymeric IgA) and had glucose concentrations less than 15 mg/dl. Fluid from both hepatic and renal cysts of subjects with autosomal dominant polycystic kidney disease, but not from the subject with the solitary hepatic cyst, demonstrated extensive changes in the electrophoretic mobility of several serum proteins. Initial intracystic pressures ranged from 16 to 40 cm H2O, were reduced 57% to 97% after aspiration of a portion of cyst fluid and were held constant during the secretion study. Within 8 min of the intravenous administration of secretin, secretion of fluid increased in two of three hepatic cysts and in the renal cyst. The electrolyte composition of cyst fluids was not altered by secretin. These data suggest that hepatic cystic epithelium has functional characteristics of biliary epithelium and that secretion by both hepatic and renal cysts may be hormonally regulated.