柯萨奇病毒IgM抗体胶体金免疫层析快速诊断试纸条的研制

目的建立一种快速检测柯萨奇病毒IgM抗体的胶体金免疫层析试纸条，并优化制备中各关键步骤的实验条件。方法以柠檬酸三钠还原法制备20nm的胶体金溶液，标记羊抗人IgM，制备免疫胶体金复合物，组装胶体金免疫层析快速诊断试纸条。结果用柠檬酸还原法制备的20nm胶体金溶液呈亮红色。胶体金标记羊抗人IgM最低稳定量是1μg/ml，最适稳定量为1．5μg/ml。最适pH为8．2。血清标本检测结果与进口ELISA试剂盒比较差异无统计学意义。结论胶体金免疫层析试纸条制备质量不但与抗原抗体的质量、层析材料的选择、胶体金的制备与标记等因素密切相关，而且缓冲系统、辅助添加剂的选择与优化也非常重要。     

免疫层析法快速测定鸡肉和鸡蛋中磺胺类药物残留

目的：用免疫层析试纸条法快速测定鸡蛋及鸡肉中磺胺类药物残留。方法：采用柠檬酸三钠还原法制备胶体金,用胶体金标记抗磺胺类药物母核结构单克隆抗体,磺胺类药物竞争物包被于硝酸纤维素膜,制成免疫层析快速检测试纸条。结果：该方法可以在15分钟内完成对磺胺类药物的半定量残留检测,检测磺胺间甲氧嘧啶（SMM）、磺胺对甲氧嘧啶（SMD）、磺胺二甲氧嘧啶（SDM）的灵敏度为10ng/ml,肉眼判定检测限为20ng/ml,样品中回收率在85%～95.6%之间;检测磺胺嘧啶（SDZ）的灵敏度为10ng/ml,肉眼判定检测限为40ng/ml,样品中的回收率在44.8%～60.9%之间。用本方法与高效液相色谱法（HPLC）对来自动物试验的鸡肉和鸡蛋样品进行对比检测,具有较高的符合率。结论：该方法灵敏度高,检测快速、准确,无需特殊仪器设备,可作为磺胺类药物残留的快速筛选方法。     

人心肌肌钙蛋白Ⅰ胶体金免疫层析试纸条的研制

目的建立一种简便实用胶体金免疫层析检测方法．用于人心肌肌钙蛋白I（cTnI）的快速检测。方法采用柠檬酸三钠还原法制备胶体金,以鼠源抗cTnI单克隆抗体进行胶体金标记,以兔源抗cTnI多克隆抗体固定于硝酸纤维素膜作为捕获抗体,采用免疫层析技术制备快速检测cThI胶体金免疫层析检测试纸条。并与国外生产的胶体金试剂盒作对比。结果该试纸条检测范围质量浓度为5ng／mL～1μg／mL;灵敏度质量浓度为5ng／mL：特异性：与肌红蛋白、肌酸磷酸激酶同工酶、cTnT、cTnC无交叉反应：与国外胶体金试纸条检测结果比较阳性符合率为96．4％,阴性符合率为100．0％。结论成功建立cTnI胶体金免疫层析检测方法,该方法灵敏度高,特异性强,检测速度快,适于急性心肌梗死的早期诊断及科研工作需要．     

快速乙肝表面抗原胶体金免疫层析测定法的建立及应用

建立乙肝表现抗原胶体金免疫层析测定法，用于乙肝病毒感染检测。采用亲和力强、特异笥高的自制抗－ＨＢｓ单克隆抗体及胶体金标记和硝酸纤维膜包被技术，制备乙肝表面抗原胶体金免疫层析快速测定试条，并进行了初步临床应用。该法的灵敏度为５ｎｇ／ｍＬ，与目前广泛使用的ＥＬＩＳＡ法对照检测５１３份血清标本，符合率为９９．２％。该法快速、准确、特异、灵敏、简便，基本符合临床应用要求，可用于乙肝表面抗原的检测。     

早期诊断心肌损伤肌球蛋白胶体金免疫层析法研究

目的建立一种简便、快速、准确检测人心肌损伤的胶体金免疫层析法（GICA）。方法制备胶体金标记抗Myosin多肽抗体，抗Myosin单克隆抗体，结合垫和样品垫的处理，组装免疫层析试纸条。检测患者血清中Myosin，进行敏感性和特异性评定。结果测试条灵敏度可达5ng／mL。检测30例急性心肌梗塞（AMI）患者血清Myosin水平，并与Roche肌钙蛋白胶体金免疫层析试剂条比较，符合率达84.2％。结论本方法特异性强、灵敏度高、简便快速、可用于急性心肌损伤的早期诊断。     

测定人绒毛膜促性腺素(HCG)的酶免疫传感器的研制

本文研制了固定HCG抗体的醋酸纤维素生物活性膜。采用夹心法在膜上发生免疫化学反应,将形成夹心结构的膜,安装在氧电极上,构成酶免疫传感器。耦联过氧化物酶的膜,催化分解H_2C_2致使传感器的输出电流提高,表示为溶液O_2含量的增高。通过溶液中O_2含量的变化值ΔC_2,测定HCG的浓度。被测HCG的浓度范围为5～2500mIU/ml。酶免疫传感器与放射免疫测定相比较,两种方法具有高度相关性,相关系数r=0.998,P<0.001。     

金标免疫层析法测定前列腺抗原方法的应用

目的：采用前列腺抗原（ＰＳＡ）单抗标记胶体金的方法。测定ＰＳＡ抗原。方法：采用柠檬酸三钠还原法制备胶体金、金标免疫层析方法（ＧＳＬＩＡ）测定。结果：将金标方法与ＥＬＩＳＡ方法进行比较灵敏度、特异性、其两者无显著性差异。金标方法可在室温放置１年以上。结论：金标免疫层析法测定前列腺抗原，是一种快速、简便、灵敏的筛选前列腺癌患者的诊断方法。     

基于CCD的胶体金免疫层析检测设备的设计

研制开发了一套用于细胞毒素相关基因A定量检测的胶体金免疫层析试纸条检测设备。该设备由硬件部分和软件系统两部分组成。硬件系统主要用于胶体金免疫层析试纸条图像的采集,而软件系统则实现了人机的交互。检测设备利用CCD(电荷耦合器件)图像传感器件采集试纸条图像,并传输至PC模块。然后,软件系统利用算法对其进行数据分析和结果诊断。软件系统主要采用了基于HSI色彩空间的移动k-均值图像分割算法,该算法实现了胶体金免疫层析试纸条图像的有效分割,完成了定量检测。本研究用10个不同浓度的胶体金免疫层析试纸条进行检测,绘制出定量曲线。还分别检测了50个阴性和50个阳性的细胞毒素相关基因A实验样品,检测结果显示特异性和灵敏度均为100%,证明了该设备的可靠性。     

两种尿hCG测定方法在临床早孕诊断中的应用评价

两种尿ｈＣＧ测定方法在临床早孕诊断中的应用评价许元初，曹燕兰，龚萍，冯颖，孙秋蓉检测尿液或血清中的ｈＣＧ对早孕诊断具有重要意义。目前最常用的方法有乳胶凝集试验、放射免疫测定、滴金免疫测定法和免疫层析试验等，我们用免疫层析试验测定尿中ｈＣＧ，该法简单方...     

制备醛固酮多克隆抗体建立检测血浆醛固酮的化学发光免疫分析法

目的：制备可用于免疫分析的醛固酮(ALD)多克隆抗体,并建立基于生物素-链霉亲和素放大系统的ALD化学发光免疫分析方法,用于测定人血液中的ALD含量。 方法：对ALD进行化学改造,制备醛固酮肟,再与BSA偶联制备免疫原,免疫兔,制备抗ALD多克隆抗体。以生物素-链霉亲和素放大系统采用竞争法建立ALD化学发光免疫分析方法。结果：经检测免疫的3号兔获得的抗ALD抗体灵敏度最高,50%抑制率（IC₅₀）ALD浓度是268 pg/ml。用该抗体建立的化学发光免疫分析法的检测范围为62.5~2 000 pg/ml,灵敏度为 23.7 pg/ml,批内变异系数为6.9%~9.5%,批间变异系数为8.5%~12.7%,回收率范围为93.1%~104.1%,稀释实验测定值与理论值呈线性相关,相关系数为r=0.996,与放射免疫分析试剂盒的相关性方程分别为y=0.932x+4.596,相关系数r=0.948（n=95）。结论：建立的检测ALD的化学发光免疫分析法符合临床应用的基本要求。     

Paired human chorionic gonadotrophin determinations for the prediction of pregnancy outcome in assisted reproduction

The aim of this study was to determine the prognostic value of single and paired measurements of serum concentrations of human chorionic gonadotrophin (HCG) for successful pregnancy following in-vitro fertilization (IVF) and tubal embryo transfer (TET). We analysed serum HCG concentrations 15 and 22 days after IVF or TET in 198 conception cycles. Cut-off values of serum HCG were determined by a receiver operating characteristic (ROC) curve. On the basis of single HCG samples on day 15 (HCG15) after transfer, using a cut-off value of HCG15 = 150 mIU/ml, the sensitivity was 71% and the specificity was 77%. The positive predictive value (HCG15 > or = 150 mIU/ml indicating a normal pregnancy) was 89%, while the negative predictive rate (HCG15 < 150 mIU/ml indicating an abnormal pregnancy) was 51%. Patients with HCG15 < 150 mIU/ml but HCG22/HCG15 ratio > or = 15, still had a 90% chance of normal pregnancy. However, in patients with HCG15 < 150 mIU/ml and an HCG22/HCG15 ratio < 15, there was an 84% chance of an abnormal pregnancy. We conclude that a single HCG15 determination combined with the ratio of HCG22 to HCG15 has a higher diagnostic accuracy for prediction of pregnancy outcome than either analysis alone.      

A sensitive kinetic latex agglutination immunoassay adapted to centrifugal analysis

Latex reagents for HCG obtained from commercially available pregnancy test kits were adapted for use on Instrumentation Laboratory's Multistat III Plus centrifugal analyzer. The clearance rate of an agglutinating reaction mixture can be measured by absorbance over a time period of 15 min. The automated system has a sensitivity of less than 5 mIU HCG/ml in buffer or urine with a range of up to 100 mIU HCG/ml. Statistical analyses on urine samples indicated CVs of between 7.6% and 10.7% for within-run precision and between 6.8% and 10.8% for between-run precision. The correlation between the centrifugal latex agglutination method and two commercially available RIAs was found to be about 87%.     

Measurement of serum chorionic gonadotropin by a biotin-avidin labeled enzyme immunoassay

A biotin-avidin enzyme immunoassay (EIA) for the measurement of human chorionic gonadotropin (hCG) in serum is described. This procedure involves the use of specific antibody immobilized on beads, biotin-labeled specific antibody, and enzyme-labeled avidin. Reproducible results were achieved within three hours for hCG in serum in the range of two mIU/ml to 150 mIU/ml. HCG levels as low as 0.4 mIU/ml can be measured. The biotin-avidin EIA and two commercially-available radioimmunoassay (RIA) kits were used to determine serum hCG levels on a group of patient samples. Good agreement was found between the biotin-avidin EIA and the RIA methods.     

血清绒毛膜促性腺激素及孕酮检测在妊娠早期中的诊断及治疗意义

目的探讨血清孕酮与绒毛膜促性腺激素联合检测在妊娠早期中的诊断及治疗意义。方法分别采用ECL／RIA技术对300例早期妊娠者血清孕酮与绒毛膜促性腺激素进行检测。结果血清P值〉60nmol／L，血β－HCG〉50mIU／ml患者85例，占28．33％，经卧床休息，未用药治疗，均胚胎发育正常；血清P值在30-60nmol／L，血β-HCG10—50／mIU／ml患者155例，占51．67％，经黄体酮与绒毛膜促性腺激素保胎治疗后均胚胎发育正常；血清P值〈30nmol／L，血β-HCG〈10mIU／ml患者45例，占15％，终止妊娠者均可见清除宫内组织物中几乎不见新鲜绒毛并伴有不同程度的陈旧性出血；血清孕酮与β-HCG上升不同步患者15例，占5．0％，均最终难免流产。结论对于妊娠早期者动态观察血清孕酮与β—HCG，对于妊娠结局的预测及指导治疗具有重要的临床价值。     

Regulation of aromatase activity in FSH-primed rat granulosa cells in vitro by follicle-stimulating hormone and various amounts of human chorionic gonadotrophin.

The influence of various amounts of human chorionic gonadotrophin (HCG), with or without follicle-stimulating hormone (FSH), on aromatase activity, progesterone- and cAMP-accumulation in rat granulosa cells was investigated. Cells were isolated from immature diethylstilbestrol-treated rats and primed for 2 days in vitro with FSH (50 mIU/ml) to give a maximum response of the cells with respect to the induction of aromatase activity. After FSH priming, small amounts of HCG (1-5 mIU/ml) increased aromatase activity during an additional culture period of 24 h, with a maximum of 4- to 5-fold at 5-8 mIU/ml HCG. At higher concentrations of HCG, aromatase activity declined reaching a plateau (1.5-fold stimulation) at 20 mIU/ml HCG. When HCG was combined with FSH (50 mIU/ml) following the priming period, aromatase activity was stimulated in an additive way, reaching a maximum at 4-5 mIU/ml HCG followed by a decline, similar to that seen in the absence of FSH. HCG induced a dose-dependent increase in cAMP with a maximum at 20 mIU/ml. This increase in cAMP was higher when HCG was combined with FSH. Aromatase activity was inhibited by 3-isobutyl-methyl-xanthine and forskolin, both of which induce elevated cAMP levels in the cell. HCG, and HCG in combination with FSH, increased the production of progesterone in a dose-dependent manner until a plateau was reached. In the presence of FSH, this plateau was reached at lower HCG concentrations. Both R5020 and progesterone showed dose-dependent inhibition of aromatase activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

HCG滴金免疫测定法试剂的研制及应用

应用自制的两种抗人绒毛膜促性腺激素（ＨＣＧ）单克隆抗体、分别包被硝酸纤维膜及制备胶体金结合物,建立了检测ＨＣＧ的滴金免疫测定法。结果表明,本法简便、快速,灵敏度高（最小抗原检测量为５０ｍＩＵ／Ｌ）,特异性好,与高浓度的ＬＨ、ＦＳＨ、ＴＳＨ均无交叉反应,临床检验２１７例样品,与ＥＬＩＳＡ法对比测定的符合率为１００％。     

Human trophectoderm biopsy and secretion of chorionic gonadotrophin.

We have previously developed a technique of trophectoderm biopsy to obtain cells from human blastocysts for preimplantation genetic diagnosis. To determine whether this technique affects the subsequent development of the blastocyst, 45 manipulated blastocysts were observed from days 3 to 14 in culture, the amount of human chorionic gonadotrophin (HCG) secreted by each embryo was measured and these results were compared with those of 26 non-manipulated controls documented in a previous study. A slit was made in the zona pellucida opposite the inner cell mass in 18 of the 45 blastocysts. This increased the rate of hatching but the other morphological changes up to day 14 were similar to those seen in the non-manipulated controls. There was no difference in the mean cumulative HCG secretion by these zona-slit controls (149.8 +/- 45.7 mIU/ml) compared to the non-manipulated controls (146.2 +/- 23.7 mIU/ml). A slit was also made in the zona pellicida of the other 27 blastocysts. Approximately 12-18 h later, in 18 blastocysts a biopsy of the herniating trophectoderm cells (5-30) was performed. The rate of hatching and adherence to the culture dish was similar to the non-manipulated controls. The mean cumulative HCG secretion decreased significantly (57.5 +/- 16.2 mIU/ml, P less than 0.01) after the biopsy procedure. However, if a small biopsy was performed (less than 10 cells removed) the decrease in HCG secretion (87.6 +/- 24.8 mIU/ml) was not significant, whereas when a large biopsy was performed (greater than 10 cells), HCG levels fell to 19.9 +/- 9.1 mIU/ml.(ABSTRACT TRUNCATED AT 250 WORDS)     

Does ICSI affect early serum beta-HCG in pregnancies achieved after IVF?

This study was conducted to compare early serum human chorionic gonadotrophin (HCG) concentrations in singleton pregnancies achieved after intracytoplasmic sperm injection (ICSI), with those achieved after conventional in-vitro fertilization (IVF). Early serum HCG, 14-16 days after embryo transfer, was analysed in 99 IVF pregnancies achieved after ICSI (group A), and compared to 105 conventional IVF pregnancies (group B). All women were treated at the IVF Unit, Lis Maternity Hospital. Records were studied retrospectively. The mean +/- SE serum HCG concentration on day 14 after embryo transfer in group A was 190.5 +/- 17.4 mIU/ml, compared to 195.7 +/- 14.03 mIU/ml in group B. HCG concentration 14 days after embryo transfer in both groups A and B was higher in women with mechanical factor than in couples with male factor infertility or unexplained infertility (246 +/- 31.4, 183.3 +/- 16.4, 177.98 +/- 14.3 mIU/ml respectively). On the 16th day after embryo transfer, the HCG concentration increased, and the difference between the groups was maintained. Only in the subgroup of unexplained infertility did we find a difference in concentrations of HCG between ICSI and conventional IVF: on the 16th day following embryo transfer in this group there was a significant difference in HCG concentrations (395. 8 +/- 21 and 545.6 +/- 45.7 respectively; P = 0.04). HCG concentrations did not differ overall in the conventional IVF pregnancies compared with those achieved by ICSI. However, a statistical difference in early serum HCG concentrations was found in relation to the aetiology of infertility.     

Enzyme-linked immunosorbent urine pregnancy tests. Clinical specificity studies

The performance of six recently introduced highly sensitive enzyme-linked immunosorbent (ELISA) urine pregnancy test reagent kits was evaluated for false positive results using 100 male and 100 postmenopausal female urine specimens and the findings were compared with those of a qualitative radioimmunoassay (RIA) procedure. Based on the findings, the most suitable pregnancy test reagent kit was then selected for doing routine pregnancy testing of urine samples of premenopausal patients. The less sensitive PregnaSTICK ELISA method and Concept RIA procedure did not give any positive results. Positive results for postmenopausal female and male urine samples were obtained as follows: Testpack, 2 and 0 (greater than 50 mIU/mL [IU/L]); Icon, 2 and 0 (greater than 50 mIU/mL [IU/L]); Quest, 4 and 2 (greater than 50 mIU/mL [IU/L]); Nimbus, 17 and 4 (greater than 25 mIU/mL [IU/L]); and Sensi-Chrome, 33 and 19 (greater than 50 mIU/mL [IU/L]), respectively. The medical records of the patients whose urine samples gave positive results were examined for information that would have explained the positive results, but no clear-cut reasons were found. Comparison of the routine urinalysis findings showed that there was a correlation between the mucus content of female (but not of male) urine samples and the incidence of false positive human chorionic gonadotropin results. During 12 months of routine use of the Icon reagents for pregnancy testing of premenopausal urine samples, the University of Texas Medical Branch staff has not reported any suspected false positive findings to the authors.