Pub Date : 2019-07-18DOI: 10.1080/15321819.2019.1642916
K. Bentum, R. Folitse, E. Amemor, V. Burimuah, T. Opoku-Agyemang, B. Emikpe
ABSTRACT Toxoplasmosis, caused by T. gondii, is an important zoonosis worldwide. In Ghana, information on the disease in humans abounds but scanty in animals. This study was therefore conducted to estimate the seroprevalence of T. gondii infection sheep and goats sampled from the Kumasi Abattoir in Ashanti Region, Ghana. A total of 347 serum samples collected from 170 sheep and 177 goats were analyzed for the presence of T. gondii antibodies using a commercial ELISA kit. Results of this study estimated the seroprevalence of 23.7% in goats an, 35.9% in sheep. In sheep, 24 (35.82%) out of a total of 67 male samples were positive and 37 (35.92%) out of a total of 104 female samples were positive while in goats, 6 (8.2%) bucks out of a total of 73 were positive while 36 (34.6%) does out of a total of 104 were positive. There was a significant difference in the rate of seropositivity of female goats (p-value 0.01). This study confirms the existence of T. gondii infection in small ruminants in Ghana and it showed that sheep and dogs are more at risk to T. gondii infection hence meat from such animals could be a potential risk to public health if consumed raw or undercooked.
{"title":"Seroprevalence of Toxoplasma Gondii antibodies in sheep and goats slaughtered at the Kumasi Abattoir, Ghana","authors":"K. Bentum, R. Folitse, E. Amemor, V. Burimuah, T. Opoku-Agyemang, B. Emikpe","doi":"10.1080/15321819.2019.1642916","DOIUrl":"https://doi.org/10.1080/15321819.2019.1642916","url":null,"abstract":"ABSTRACT Toxoplasmosis, caused by T. gondii, is an important zoonosis worldwide. In Ghana, information on the disease in humans abounds but scanty in animals. This study was therefore conducted to estimate the seroprevalence of T. gondii infection sheep and goats sampled from the Kumasi Abattoir in Ashanti Region, Ghana. A total of 347 serum samples collected from 170 sheep and 177 goats were analyzed for the presence of T. gondii antibodies using a commercial ELISA kit. Results of this study estimated the seroprevalence of 23.7% in goats an, 35.9% in sheep. In sheep, 24 (35.82%) out of a total of 67 male samples were positive and 37 (35.92%) out of a total of 104 female samples were positive while in goats, 6 (8.2%) bucks out of a total of 73 were positive while 36 (34.6%) does out of a total of 104 were positive. There was a significant difference in the rate of seropositivity of female goats (p-value 0.01). This study confirms the existence of T. gondii infection in small ruminants in Ghana and it showed that sheep and dogs are more at risk to T. gondii infection hence meat from such animals could be a potential risk to public health if consumed raw or undercooked.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"25 1","pages":"495 - 501"},"PeriodicalIF":0.0,"publicationDate":"2019-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86843441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1250772
A. Rodríguez-Rodríguez, J. Egea-Guerrero, Á. Vilches-Arenas, Manuel Jesús Quintanilla-Vázquez, F. Murillo-Cabezas, M. Muñoz-Sanchez
ABSTRACT Changes in Urotensin-II (U-II) concentration, a potent vasoconstrictor peptide, have been detected in various pathologies, but it has been impossible to define a normality range. We aimed to analyze the concordance and interchangeability between two enzyme immunoassay methods developed by Phoenix Pharmaceuticals, Inc. to measure U-II plasma concentration in rats: ELISA and fluorescent EIA. Assays resulted positively correlated (r = 0.850; p < 0.01). There was a significant difference between assays values (p < 0.001). The analysis of agreement (Bland and Altman plot) stated that the mean of the differences was 2.055 (SD ± 0.588). Hence, we concluded that the two U-II assays were correlated but not interchangeable.
{"title":"Comparison of two competitive enzyme immunoassay kits for quantification of plasma Urotensin-II in rats","authors":"A. Rodríguez-Rodríguez, J. Egea-Guerrero, Á. Vilches-Arenas, Manuel Jesús Quintanilla-Vázquez, F. Murillo-Cabezas, M. Muñoz-Sanchez","doi":"10.1080/15321819.2016.1250772","DOIUrl":"https://doi.org/10.1080/15321819.2016.1250772","url":null,"abstract":"ABSTRACT Changes in Urotensin-II (U-II) concentration, a potent vasoconstrictor peptide, have been detected in various pathologies, but it has been impossible to define a normality range. We aimed to analyze the concordance and interchangeability between two enzyme immunoassay methods developed by Phoenix Pharmaceuticals, Inc. to measure U-II plasma concentration in rats: ELISA and fluorescent EIA. Assays resulted positively correlated (r = 0.850; p < 0.01). There was a significant difference between assays values (p < 0.001). The analysis of agreement (Bland and Altman plot) stated that the mean of the differences was 2.055 (SD ± 0.588). Hence, we concluded that the two U-II assays were correlated but not interchangeable.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"47 1","pages":"247 - 256"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86036175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1250771
B. Jain, J. Kumarasamy, C. Gholve, S. Kulkarni, M. Rajan
ABSTRACT We describe the development and validation of multianalyte immunoassays (MAIA) for three analytes, viz., thyroxine (T4), thyroid stimulating hormone (TSH), and thyroglobulin (Tg) essential for assessment of thyroid function but having widely varying molecular weights. Using polycarbonate (PC) track-etched membranes (TEM) as an immobilization support and 125I as the tracer, both competitive assay for T4 and non-competitive assay for TSH and Tg were performed on the same TEM. MAIA was found to be highly sensitive and precise with clinically useful working range and correlated very well with individual analyte immunoassays. While we have demonstrated this assay format with radiotracer, it can be used with non-isotopic tracers equally well.
{"title":"A multi-analyte immunoassay for thyroid related analytes","authors":"B. Jain, J. Kumarasamy, C. Gholve, S. Kulkarni, M. Rajan","doi":"10.1080/15321819.2016.1250771","DOIUrl":"https://doi.org/10.1080/15321819.2016.1250771","url":null,"abstract":"ABSTRACT We describe the development and validation of multianalyte immunoassays (MAIA) for three analytes, viz., thyroxine (T4), thyroid stimulating hormone (TSH), and thyroglobulin (Tg) essential for assessment of thyroid function but having widely varying molecular weights. Using polycarbonate (PC) track-etched membranes (TEM) as an immobilization support and 125I as the tracer, both competitive assay for T4 and non-competitive assay for TSH and Tg were performed on the same TEM. MAIA was found to be highly sensitive and precise with clinically useful working range and correlated very well with individual analyte immunoassays. While we have demonstrated this assay format with radiotracer, it can be used with non-isotopic tracers equally well.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"18 1","pages":"271 - 284"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74437205","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1250774
Maysam Mard-Soltani, M. Rasaee, A. Sheikhi, M. Hedayati
ABSTRACT Designing novel antigens to rise specific antibodies for Thyroid Stimulating Hormone (TSH) detection is of great significance. A novel fusion protein consisting of the C termini sequence of TSH beta subunit and a fusion sequence was designed and produced for rabbit immunization. Thereafter, the produced antibodies were purified and characterized for TSH detection. Our results indicate that the produced antibody is capable of sensitive and specific detection of TSH with low cross reactivity. This study underscores the applicability of designed fusion protein for specific and sensitive polyclonal antibody production and the importance of selecting an amenable region of the TSH for immunization.
{"title":"Eliciting an antibody response against a recombinant TSH containing fusion protein","authors":"Maysam Mard-Soltani, M. Rasaee, A. Sheikhi, M. Hedayati","doi":"10.1080/15321819.2016.1250774","DOIUrl":"https://doi.org/10.1080/15321819.2016.1250774","url":null,"abstract":"ABSTRACT Designing novel antigens to rise specific antibodies for Thyroid Stimulating Hormone (TSH) detection is of great significance. A novel fusion protein consisting of the C termini sequence of TSH beta subunit and a fusion sequence was designed and produced for rabbit immunization. Thereafter, the produced antibodies were purified and characterized for TSH detection. Our results indicate that the produced antibody is capable of sensitive and specific detection of TSH with low cross reactivity. This study underscores the applicability of designed fusion protein for specific and sensitive polyclonal antibody production and the importance of selecting an amenable region of the TSH for immunization.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"13 1","pages":"257 - 270"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90537121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1260587
A. Roșca, G. Anton, L. Ene, I. Iancu, A. Temereanca, C. Achim, S. Ruță
ABSTRACT This study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p = 0.01) and DNMT3A (p = 0.004) independently correlated with active viral replication. DNMT3A expression increased with total treatment duration (p = 0.03), number of antiretroviral drugs ever used (p = 0.003), and cumulative exposure to protease inhibitors (p = 0.02) even in currently HIV undetectable patients.
{"title":"Immunoassay and molecular methods to investigate DNA methylation changes in peripheral blood mononuclear cells in HIV infected patients on cART","authors":"A. Roșca, G. Anton, L. Ene, I. Iancu, A. Temereanca, C. Achim, S. Ruță","doi":"10.1080/15321819.2016.1260587","DOIUrl":"https://doi.org/10.1080/15321819.2016.1260587","url":null,"abstract":"ABSTRACT This study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p = 0.01) and DNMT3A (p = 0.004) independently correlated with active viral replication. DNMT3A expression increased with total treatment duration (p = 0.03), number of antiretroviral drugs ever used (p = 0.003), and cumulative exposure to protease inhibitors (p = 0.02) even in currently HIV undetectable patients.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"116 1","pages":"299 - 307"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89323965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1260584
Susanne Pribbenow, T. G. Shrivastav, M. Dehnhard
ABSTRACT Enzyme-immunoassays (EIA) that detect fecal testosterone metabolites (fTM) are powerful tools to monitor gonadal activity non-invasively. However, a challenge with testosterone EIAs might be their potential for cross-reactivities with structurally similar glucocorticoid metabolites. Therefore, we aimed to verify the capability of four different testosterone EIAs to monitor fTM without reflecting changes in adrenocortical activity in spotted hyenas by analyzing fecal samples following testosterone and ACTH challenge tests. We demonstrated that none of the testosterone EIAs is appropriate to measure fTM as all of them showed substantial cross-reactivities to unknown metabolites. Our study underlines the importance of validating androgen EIAs.
{"title":"Measuring fecal testosterone metabolites in spotted hyenas: Choosing the wrong assay may lead to erroneous results","authors":"Susanne Pribbenow, T. G. Shrivastav, M. Dehnhard","doi":"10.1080/15321819.2016.1260584","DOIUrl":"https://doi.org/10.1080/15321819.2016.1260584","url":null,"abstract":"ABSTRACT Enzyme-immunoassays (EIA) that detect fecal testosterone metabolites (fTM) are powerful tools to monitor gonadal activity non-invasively. However, a challenge with testosterone EIAs might be their potential for cross-reactivities with structurally similar glucocorticoid metabolites. Therefore, we aimed to verify the capability of four different testosterone EIAs to monitor fTM without reflecting changes in adrenocortical activity in spotted hyenas by analyzing fecal samples following testosterone and ACTH challenge tests. We demonstrated that none of the testosterone EIAs is appropriate to measure fTM as all of them showed substantial cross-reactivities to unknown metabolites. Our study underlines the importance of validating androgen EIAs.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"50 1","pages":"308 - 321"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89149967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-05-04DOI: 10.1080/15321819.2016.1266499
Ahad Khalilnezhad, Elham Mahmoudian, N. Mosaffa, J. Mohsenifar, D. Amani
ABSTRACT Background. We hypothesized that Tumor cell lysate (TCL) prepared from spontaneous mouse mammary tumor (SMMT) may elicit IgG production by spleen mononuclear cells (SMCs) in ex vivo. Methods. The SMCs from healthy mice (HM, n = 6) and four-week SMMT-bearing mice (TBM, n = 6) was cultured in presence of TCL and mitogen for 42 hr at 37°C, separately. Serum and SMCs culture supernatant levels of IFNγ, IL-4, and total IgG were measured using special ELISA kits. Results. Serum IgG level of TBM was significantly higher than that of HM group (P = 0.019), while serum IFNγ and IL-4 did not differ between two groups (P > 0.05). Mitogen significantly induced ex vivo production of both IFNγ (P = 0.013) and IL-4 (P = 0.015) by SMCs from HM group, and only IL-4 (P = 0.049) by SMCs from TBM group. In contrast, TCL increased ex vivo production of IgG by SMCs from both HM (P = 0.034) and TBM (P = 0.016) groups. The ex vivo IgG revealed a moderate positive correlation with tumor size (r = 0.578, P = 0.422). Conclusion. It seems that TCL prepared from SMMT are potent inducers of IgG production. This may propose TCL as a potential tool for monitoring of humoral immunity in animal models.
{"title":"Spontaneous mouse mammary tumor cell lysates induce IgG production in spleen mononuclear cells of healthy and tumor-bearing mice","authors":"Ahad Khalilnezhad, Elham Mahmoudian, N. Mosaffa, J. Mohsenifar, D. Amani","doi":"10.1080/15321819.2016.1266499","DOIUrl":"https://doi.org/10.1080/15321819.2016.1266499","url":null,"abstract":"ABSTRACT Background. We hypothesized that Tumor cell lysate (TCL) prepared from spontaneous mouse mammary tumor (SMMT) may elicit IgG production by spleen mononuclear cells (SMCs) in ex vivo. Methods. The SMCs from healthy mice (HM, n = 6) and four-week SMMT-bearing mice (TBM, n = 6) was cultured in presence of TCL and mitogen for 42 hr at 37°C, separately. Serum and SMCs culture supernatant levels of IFNγ, IL-4, and total IgG were measured using special ELISA kits. Results. Serum IgG level of TBM was significantly higher than that of HM group (P = 0.019), while serum IFNγ and IL-4 did not differ between two groups (P > 0.05). Mitogen significantly induced ex vivo production of both IFNγ (P = 0.013) and IL-4 (P = 0.015) by SMCs from HM group, and only IL-4 (P = 0.049) by SMCs from TBM group. In contrast, TCL increased ex vivo production of IgG by SMCs from both HM (P = 0.034) and TBM (P = 0.016) groups. The ex vivo IgG revealed a moderate positive correlation with tumor size (r = 0.578, P = 0.422). Conclusion. It seems that TCL prepared from SMMT are potent inducers of IgG production. This may propose TCL as a potential tool for monitoring of humoral immunity in animal models.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"1 1","pages":"333 - 342"},"PeriodicalIF":0.0,"publicationDate":"2017-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82436208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-03-04DOI: 10.1080/15321819.2016.1234485
K. Kerboua, F. Haiba, D. Batouche
ABSTRACT Treatment of atypical hemolytic uremic syndrome (aHUS) by the complement C5 inhibitor eculizumab (Soliris®) is highly effective but unfortunately, associated with an economic pressure on the health care systems even in high incomes countries. Despite spacing infusions having been proposed as the unique solution to minimize this economic impact, no reliable laboratory assays are available to tailor such therapy optimization. We aimed to propose and evaluate a complement composite marker for eculizumab efficacy monitoring in aHUS. We have retrospectively analyzed complement profiles data of eight aHUS patients under eculizumab from the International Registry of HUS/Thrombotic Thrombocytopenia Purpura, and calculated a novel marker “C3:CH50 ratio” by dividing C3 value by CH50 one for each sample during induction and maintenance periods. The results significance was compared to the currently used biomarkers for eculizumab tailoring. In contrast to the current biomarkers used for eculizumab efficacy monitoring like CH50 and soluble or deposit membrane attack complexes, “C3:CH50 ratio” seems to be the most interesting one since its value at pre-eculizumab dosage equaled 0.92 ± 0.2 while the post-eculizumab one increased significantly to reach 24.54 ± 10.7; P < 0.001. Furthermore, this ratio correlated negatively with platelets count (r = −0.722, P = 0.018) while no correlation was found within the thrombotic microangiopathy (TMA) biomarkers and complement blockade for the other parameters that change in pre and post-eculizumab therapy. As far as we know, this is the first study that suggests a post-eculizumab parameter correlating simultaneously with drug’s activity (complement inhibition) and disease activity (platelets counts). Nonetheless, the limited number of patients enrolled in this study should be considered in larger studies to guide eculizumab optimization by indicating the time when subsequent withdrawal or infusion spacing is allowed or recommended.
补体C5抑制剂eculizumab (Soliris®)治疗非典型溶血性尿毒症综合征(aHUS)非常有效，但不幸的是，即使在高收入国家，也与卫生保健系统的经济压力相关。尽管间隔输注被认为是减少这种经济影响的唯一解决方案，但目前还没有可靠的实验室分析方法来定制这种疗法的优化。我们的目的是提出并评估一种补体复合标记物，用于监测eculizumab在aHUS中的疗效。我们回顾性分析了来自国际溶血性尿毒综合征/血栓性血小板减少性紫癜登记处的8例eculizumab下的aHUS患者的补体谱数据，并通过在诱导和维持期间将每个样本的C3值除以CH50，计算出一个新的标记物“C3:CH50比率”。结果的意义与目前用于eculizumab裁剪的生物标志物进行了比较。与目前用于eculizumab疗效监测的生物标志物(如CH50和可溶性或沉积膜攻击复合物)相比，“C3:CH50比值”似乎是最有趣的，因为在eculizumab给药前其值为0.92±0.2，而eculizumab给药后其值显著增加，达到24.54±10.7;P < 0.001。此外，该比值与血小板计数呈负相关(r = - 0.722, P = 0.018)，而血栓性微血管病变(TMA)生物标志物和补体阻断在eculizumab治疗前后的其他参数变化中未发现相关性。据我们所知，这是第一个表明eculizumab后参数与药物活性(补体抑制)和疾病活性(血小板计数)同时相关的研究。尽管如此，在更大规模的研究中，应该考虑纳入本研究的有限患者数量，通过指示允许或推荐后续停药或输液间隔的时间来指导eculizumab优化。
{"title":"C3:CH50 ratio as a proposed composite marker for eculizumab monitoring in atypical hemolytic uremic syndrome: Preliminary results","authors":"K. Kerboua, F. Haiba, D. Batouche","doi":"10.1080/15321819.2016.1234485","DOIUrl":"https://doi.org/10.1080/15321819.2016.1234485","url":null,"abstract":"ABSTRACT Treatment of atypical hemolytic uremic syndrome (aHUS) by the complement C5 inhibitor eculizumab (Soliris®) is highly effective but unfortunately, associated with an economic pressure on the health care systems even in high incomes countries. Despite spacing infusions having been proposed as the unique solution to minimize this economic impact, no reliable laboratory assays are available to tailor such therapy optimization. We aimed to propose and evaluate a complement composite marker for eculizumab efficacy monitoring in aHUS. We have retrospectively analyzed complement profiles data of eight aHUS patients under eculizumab from the International Registry of HUS/Thrombotic Thrombocytopenia Purpura, and calculated a novel marker “C3:CH50 ratio” by dividing C3 value by CH50 one for each sample during induction and maintenance periods. The results significance was compared to the currently used biomarkers for eculizumab tailoring. In contrast to the current biomarkers used for eculizumab efficacy monitoring like CH50 and soluble or deposit membrane attack complexes, “C3:CH50 ratio” seems to be the most interesting one since its value at pre-eculizumab dosage equaled 0.92 ± 0.2 while the post-eculizumab one increased significantly to reach 24.54 ± 10.7; P < 0.001. Furthermore, this ratio correlated negatively with platelets count (r = −0.722, P = 0.018) while no correlation was found within the thrombotic microangiopathy (TMA) biomarkers and complement blockade for the other parameters that change in pre and post-eculizumab therapy. As far as we know, this is the first study that suggests a post-eculizumab parameter correlating simultaneously with drug’s activity (complement inhibition) and disease activity (platelets counts). Nonetheless, the limited number of patients enrolled in this study should be considered in larger studies to guide eculizumab optimization by indicating the time when subsequent withdrawal or infusion spacing is allowed or recommended.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"29 1","pages":"178 - 189"},"PeriodicalIF":0.0,"publicationDate":"2017-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73706426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-03-04DOI: 10.1080/15321819.2016.1236729
Elisa M. Castells Martínez, Ruben Del Valle, E. C. González, A. Melchor, P. L. Pérez, Idania González, A. Carr, K. Leon
ABSTRACT Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay. The EGF assay was completed in 18 hr, with a measuring range of 39–2500 pg/mL. The intra- and inter-assay coefficients of variation were 4.4–7.3% and 0–5.1%, respectively, depending on the EGF concentrations evaluated. Percentage recovery ranged from 96–104%. Regression analysis showed a good correlation with the commercially available Human EGF Immunoassay Quantikine® ELISA kit (n = 130, r = 0.92, P < 0.01). The analytical performance characteristics of our UMELISA EGF endorse its use for the quantification of EGF in human serum samples.
人表皮生长因子是一种由53个氨基酸残基组成的小肽，它能刺激细胞增殖，并与几种人类癌症有关。利用链霉亲和素与生物素之间高亲和力反应的优点，建立了一种简便的三明治型超微elisa (UMELISA)检测人血清中EGF的方法。用高亲和力单克隆抗体包被的条带作为固相，以确保检测的特异性。EGF测定在18小时内完成，测量范围为39-2500 pg/mL。根据评估的EGF浓度，测定内和测定间的变异系数分别为4.4-7.3%和0-5.1%。回收率为96 ~ 104%。回归分析显示，与市售的Human EGF Immunoassay Quantikine®ELISA试剂盒相关性良好(n = 130, r = 0.92, P < 0.01)。我们的UMELISA EGF的分析性能特点支持其用于人血清样品中EGF的定量。
{"title":"An enzyme immunoassay for determining epidermal growth factor (EGF) in human serum samples using an ultramicroanalytical system","authors":"Elisa M. Castells Martínez, Ruben Del Valle, E. C. González, A. Melchor, P. L. Pérez, Idania González, A. Carr, K. Leon","doi":"10.1080/15321819.2016.1236729","DOIUrl":"https://doi.org/10.1080/15321819.2016.1236729","url":null,"abstract":"ABSTRACT Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay. The EGF assay was completed in 18 hr, with a measuring range of 39–2500 pg/mL. The intra- and inter-assay coefficients of variation were 4.4–7.3% and 0–5.1%, respectively, depending on the EGF concentrations evaluated. Percentage recovery ranged from 96–104%. Regression analysis showed a good correlation with the commercially available Human EGF Immunoassay Quantikine® ELISA kit (n = 130, r = 0.92, P < 0.01). The analytical performance characteristics of our UMELISA EGF endorse its use for the quantification of EGF in human serum samples.","PeriodicalId":15987,"journal":{"name":"Journal of Immunoassay and Immunochemistry","volume":"1 1","pages":"190 - 201"},"PeriodicalIF":0.0,"publicationDate":"2017-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89948991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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