Chronic hepatitis C virus (HCV) related liver diseases are still an ongoing cause of hepatic failure despite the effective role of direct-acting anti-viral agents. Farnesoid X receptor (FXR) agonists have a potential therapeutic effect on the management of chronic liver diseases (CLD). However, data regarding FXR protein expression in human CLDs are limited and conflicting. We aimed to assess the immunohistochemical expression of FXR in HCV-related chronic hepatitis and cirrhosis in comparison with metabolic-associated fatty liver disease (MAFLD) and normal liver tissue. The expression of FXR was low both in hepatocytes and bile ducts of HCV-related chronic hepatitis and cirrhosis (p = .001, respectively). In addition, a significantly low expression of FXR was observed in HCV-related hepatitis and cirrhosis groups compared to MAFLD in hepatocytes (p < .001, for both) and bile ducts (p = .004 and p = .018). FXR expression in HCV-related cirrhosis was significantly associated with compensated liver function (p = .032) and low inflammatory activity (p = .022). FXR expression decreases in HCV-related CLDs. There was some evidence that FXR expression could protect against post-hepatitis cirrhosis.
{"title":"Farnesoid X receptor in chronic liver diseases: an immunohistochemical study.","authors":"Dina Sweed, Nanis Shawky Holah, Esraa Karman, Nancy Asaad, Shereen Mahmoud","doi":"10.1080/15321819.2023.2242920","DOIUrl":"10.1080/15321819.2023.2242920","url":null,"abstract":"<p><p>Chronic hepatitis C virus (HCV) related liver diseases are still an ongoing cause of hepatic failure despite the effective role of direct-acting anti-viral agents. Farnesoid X receptor (FXR) agonists have a potential therapeutic effect on the management of chronic liver diseases (CLD). However, data regarding FXR protein expression in human CLDs are limited and conflicting. We aimed to assess the immunohistochemical expression of FXR in HCV-related chronic hepatitis and cirrhosis in comparison with metabolic-associated fatty liver disease (MAFLD) and normal liver tissue. The expression of FXR was low both in hepatocytes and bile ducts of HCV-related chronic hepatitis and cirrhosis (<i>p</i> = .001, respectively). In addition, a significantly low expression of FXR was observed in HCV-related hepatitis and cirrhosis groups compared to MAFLD in hepatocytes (<i>p</i> < .001, for both) and bile ducts (<i>p</i> = .004 and <i>p</i> = .018). FXR expression in HCV-related cirrhosis was significantly associated with compensated liver function (<i>p</i> = .032) and low inflammatory activity (<i>p</i> = .022). FXR expression decreases in HCV-related CLDs. There was some evidence that FXR expression could protect against post-hepatitis cirrhosis.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":" ","pages":"381-395"},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10519920","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 : 2023-11-02Epub Date: 2023-10-31DOI: 10.1080/15321819.2023.2259454
Kiana Ketabi, Hoorieh Soleimanjahi, Ala Habibian, Saied Abroun
The SARS-CoV-2 outbreak led to a health crisis worldwide. This infection can infect individuals, particularly pregnant women. In this review, we tried to find the possibility of vertical transmission of COVID-19 and investigate the effects of COVID-19 on pregnancy, breastfeeding, cord blood banking, and the effects of recommended vaccines on pregnant and lactating women. Keywords include COVID-19, congenital infection, SARS-CoV-2, pregnancy, and COVID-19 vaccines. Vertical transmission of SARS-CoV-2 was searched in scientific databases, such as PubMed, Google Scholar, and Scopus. The criteria for including studies in this article are the study of SARS-CoV-2 infection in pregnant women, fetuses, and neonates during pregnancy and while breastfeeding, and also the effect of COVID-19 vaccines on them. There are several conflicting results in the transmission of SARS-CoV-2 from the maternal-fetal interface. Since many neonates born from COVID-19-infected mothers had no signs of this infection, the possibility of SARS-CoV-2 congenital transmission cannot be confirmed. Also, SARS-CoV-2-infected women can breastfeed their babies if they have mild symptoms. Up till now, no adverse effect of COVID-19 vaccines has been identified on mothers, infants, and the fertility of men or women. Even so, more investigations are needed on the long-term effects of COVID-19 vaccines.
{"title":"Pregnancy and <i>SARS-CoV-2</i> infection with a focus on its vertical transmission, breastfeeding, cord blood banking, and vaccination during COVID-19 infection.","authors":"Kiana Ketabi, Hoorieh Soleimanjahi, Ala Habibian, Saied Abroun","doi":"10.1080/15321819.2023.2259454","DOIUrl":"10.1080/15321819.2023.2259454","url":null,"abstract":"<p><p>The <i>SARS-CoV-2</i> outbreak led to a health crisis worldwide. This infection can infect individuals, particularly pregnant women. In this review, we tried to find the possibility of vertical transmission of COVID-19 and investigate the effects of COVID-19 on pregnancy, breastfeeding, cord blood banking, and the effects of recommended vaccines on pregnant and lactating women. Keywords include COVID-19, congenital infection, <i>SARS-CoV-2</i>, pregnancy, and COVID-19 vaccines. Vertical transmission of <i>SARS-CoV-2</i> was searched in scientific databases, such as PubMed, Google Scholar, and Scopus. The criteria for including studies in this article are the study of <i>SARS-CoV-2</i> infection in pregnant women, fetuses, and neonates during pregnancy and while breastfeeding, and also the effect of COVID-19 vaccines on them. There are several conflicting results in the transmission of <i>SARS-CoV-2</i> from the maternal-fetal interface. Since many neonates born from COVID-19-infected mothers had no signs of this infection, the possibility of <i>SARS-CoV-2</i> congenital transmission cannot be confirmed. Also, <i>SARS-CoV-2</i>-infected women can breastfeed their babies if they have mild symptoms. Up till now, no adverse effect of COVID-19 vaccines has been identified on mothers, infants, and the fertility of men or women. Even so, more investigations are needed on the long-term effects of COVID-19 vaccines.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":" ","pages":"361-380"},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41124932","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 : 2023-11-02Epub Date: 2023-10-31DOI: 10.1080/15321819.2023.2260480
Moumita Paul, Bibhuti Saha, Sumi Mukhopadhyay
Rapid diagnosis of patients with severe Dengue infection can be useful for the efficient clinical management of cases caused by the Dengue virus. Lateral Flow Immunoassay (LFIA) have been broadly used for rapid Dengue diagnosis, because of their quick readouts with the human eye, simplicity of use, and affordability. Despite the availability of several commercial Dengue point-of-care assays, none has shown to be successful in discriminating between severe and nonsevere forms of Dengue infection. In the current study, for the first time, a novel lectin-based point-of-care assay for the early detection of patients with severe Dengue infection with gold-adorned sheets as detection labels is being reported. In this assay, Dengue severity was diagnosed by detecting the glycosylation profile of vitronectin, a known Dengue severity marker. Two lectins were employed namely DSA (Datura stramonium) and MAA (Maackia amurensis) that can recognize specific glycans like galactose Gal-(1-4) GlcNAc and sialic acid in an (α2-3) linkage, which displayed high sensitivity and high specificity, i.e. 90% and 85% for DSA and 90.91% and 95% for MAA. The new assay has a detection limit of 5 µg µl-1 and enables the quick (30 min) and sensitive detection of severe Dengue cases. The reported point-of-care immunoassay exhibits considerable promise for early identification of patients with Dengue severity.
{"title":"Development of a novel lectin-based gold nanoparticle point-of-care immunoassay for rapid diagnosis of patients with severe Dengue infection.","authors":"Moumita Paul, Bibhuti Saha, Sumi Mukhopadhyay","doi":"10.1080/15321819.2023.2260480","DOIUrl":"10.1080/15321819.2023.2260480","url":null,"abstract":"<p><p>Rapid diagnosis of patients with severe Dengue infection can be useful for the efficient clinical management of cases caused by the Dengue virus. Lateral Flow Immunoassay (LFIA) have been broadly used for rapid Dengue diagnosis, because of their quick readouts with the human eye, simplicity of use, and affordability. Despite the availability of several commercial Dengue point-of-care assays, none has shown to be successful in discriminating between severe and nonsevere forms of Dengue infection. In the current study, for the first time, a novel lectin-based point-of-care assay for the early detection of patients with severe Dengue infection with gold-adorned sheets as detection labels is being reported. In this assay, Dengue severity was diagnosed by detecting the glycosylation profile of vitronectin, a known Dengue severity marker. Two lectins were employed namely DSA (<i>Datura stramonium</i>) and MAA (<i>Maackia amurensis</i>) that can recognize specific glycans like galactose Gal-(1-4) GlcNAc and sialic acid in an (α2-3) linkage, which displayed high sensitivity and high specificity, i.e. 90% and 85% for DSA and 90.91% and 95% for MAA. The new assay has a detection limit of 5 µg µl<sup>-1</sup> and enables the quick (30 min) and sensitive detection of severe Dengue cases. The reported point-of-care immunoassay exhibits considerable promise for early identification of patients with Dengue severity.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":" ","pages":"418-435"},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41131076","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 : 2023-11-02Epub Date: 2023-09-11DOI: 10.1080/15321819.2023.2251564
Noha Elkady, Amira I Aldesoky, Dina Mohamed Allam
Gastric carcinoma (GC) is one of the most prevalent cancers worldwide and the fourth leading cause of cancer-related death. Studying the molecular profile of GC is essential for developing targeted therapies. β-catenin, Tenascin, and Fascin expression are among the molecular abnormalities that are claimed to cause GC progression and chemoresistance. Therefore, they could be used as potential therapeutic targets. This study aimed to evaluate β-catenin, Tenascin, and Fascin expression and their possible roles as prognostic and predictive biomarkers in GC using immunohistochemistry. This retrospective study included 84 GC cases. Tissue microarrays were constructed, followed by β-catenin, Tenascin, and Fascin immunostaining. Their expression was assessed and compared with clinicopathological parameters and survival data. The study results revealed that β-catenin nucleocytoplasmic expression, positive Tenascin, and Fascin expressions were detected in 86.9%, 70%, and 59.5% of cases, respectively. Their expression was significantly associated with poor prognostic parameters, such as deeper tumor invasion, lymph node metastasis, advanced pathological stage, vascular invasion, positive omental nodules, poor response to chemotherapy, and short overall survival. Hence, nucleocytoplasmic β-catenin expression together with Tenascin and Fascin positivity can be potential prognostic and predictive markers, and they can be used as therapeutic targets for GC.
{"title":"Can β-catenin, Tenascin and Fascin be potential biomarkers for personalized therapy in Gastric carcinoma?","authors":"Noha Elkady, Amira I Aldesoky, Dina Mohamed Allam","doi":"10.1080/15321819.2023.2251564","DOIUrl":"10.1080/15321819.2023.2251564","url":null,"abstract":"<p><p>Gastric carcinoma (GC) is one of the most prevalent cancers worldwide and the fourth leading cause of cancer-related death. Studying the molecular profile of GC is essential for developing targeted therapies. β-catenin, Tenascin, and Fascin expression are among the molecular abnormalities that are claimed to cause GC progression and chemoresistance. Therefore, they could be used as potential therapeutic targets. This study aimed to evaluate β-catenin, Tenascin, and Fascin expression and their possible roles as prognostic and predictive biomarkers in GC using immunohistochemistry. This retrospective study included 84 GC cases. Tissue microarrays were constructed, followed by β-catenin, Tenascin, and Fascin immunostaining. Their expression was assessed and compared with clinicopathological parameters and survival data. The study results revealed that β-catenin nucleocytoplasmic expression, positive Tenascin, and Fascin expressions were detected in 86.9%, 70%, and 59.5% of cases, respectively. Their expression was significantly associated with poor prognostic parameters, such as deeper tumor invasion, lymph node metastasis, advanced pathological stage, vascular invasion, positive omental nodules, poor response to chemotherapy, and short overall survival. Hence, nucleocytoplasmic β-catenin expression together with Tenascin and Fascin positivity can be potential prognostic and predictive markers, and they can be used as therapeutic targets for GC.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":" ","pages":"396-417"},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10202835","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 : 2023-07-04DOI: 10.1080/15321819.2023.2232507
{"title":"Statement of Retraction: Elevated CSF concentration of CCL3 and CCL4 in relapsing remitting multiple sclerosis patients.","authors":"","doi":"10.1080/15321819.2023.2232507","DOIUrl":"https://doi.org/10.1080/15321819.2023.2232507","url":null,"abstract":"","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"44 4","pages":"360"},"PeriodicalIF":0.0,"publicationDate":"2023-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9843484","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}
Studying the expression of hematopoietic stem cell markers from different sources might be useful in understanding stem cell biology in different niche conditions. The study aimed to assess the difference in cell surface markers (CD44, CD90, CD96) on hematopoietic stem cells in three different niche conditions; umbilical cord blood (UCB), normal bone marrow (NBM) and bone marrow samples from idiopathic (immune) thrombocytopenic purpura (IBM). This study was conducted on 300 cases divided into three study groups; 100 umbilical cord blood units collected from mothers undergoing cesarian section in gynecology and obstetrics department, 100 bone marrow samples from idiopathic (immune) thrombocytopenic purpura patients collected from university children hospital and 100 normal bone marrow samples with no evidence of disease in bone marrow tissue. CD44 was significantly elevated in UCB and NBM groups compared to IBM group (<0.001). There was also a significant elevation of CD90 and CD96 in IBM group compared to NBM group and UCB (<0.001). CD90 and CD96 play a role in the pathogenesis of ITP disorder and could be applied as a targeted therapy to improve the outcome of this disease.
{"title":"CD44, CD90 and CD96 expression in immune thrombocytopenia purpura (ITP) patients.","authors":"Nadia ElMenshawy, Farha El-Chennawi, Ahmed Darwish, Asmaa Foda, Doaa Atita, Mohamed Eissa","doi":"10.1080/15321819.2023.2191259","DOIUrl":"https://doi.org/10.1080/15321819.2023.2191259","url":null,"abstract":"<p><p>Studying the expression of hematopoietic stem cell markers from different sources might be useful in understanding stem cell biology in different niche conditions. The study aimed to assess the difference in cell surface markers (CD44, CD90, CD96) on hematopoietic stem cells in three different niche conditions; umbilical cord blood (UCB), normal bone marrow (NBM) and bone marrow samples from idiopathic (immune) thrombocytopenic purpura (IBM). This study was conducted on 300 cases divided into three study groups; 100 umbilical cord blood units collected from mothers undergoing cesarian section in gynecology and obstetrics department, 100 bone marrow samples from idiopathic (immune) thrombocytopenic purpura patients collected from university children hospital and 100 normal bone marrow samples with no evidence of disease in bone marrow tissue. CD44 was significantly elevated in UCB and NBM groups compared to IBM group (<0.001). There was also a significant elevation of CD90 and CD96 in IBM group compared to NBM group and UCB (<0.001). CD90 and CD96 play a role in the pathogenesis of ITP disorder and could be applied as a targeted therapy to improve the outcome of this disease.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"44 4","pages":"326-337"},"PeriodicalIF":0.0,"publicationDate":"2023-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9891516","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}
Immunoassay tests are used in clinical and forensic toxicology laboratories to determine illicit drug use in biological samples. Therefore, this study aims to optimize the cutoff concentrations of DOA I Plus in the blood and compare the LC-MS/MS results. 680 authentic forensic whole blood specimens with Randox Evidence DOA I Plus array were screened for drug of abuse and confirmed by LC-MS/MS. Regarding the manufacturer recommended threshold values, 139 out of 680 authentic blood samples were positive for one or more analytes, while 541 were negative. Nearly all of the 139 positive blood samples confirmed by LC-MS/MS were true positive for one or more analytes, while 522 of the 541 negative blood samples were true negative. The overall sensitivity and the specificity were 87.8% and 99.6%, respectively. THC was considered in detail, and a receiver operator characteristic curve analysis was performed to determine the optimum cutoff for THC, as it accounts for 78% of all positive results according to the manufacturer's recommended thresholds. The optimal threshold value for THC was determined at a concentration of 23 ng/mL, while these values for other parameters were defined as recommended by the manufacturer.
免疫分析测试用于临床和法医毒理学实验室,以确定生物样本中的非法药物使用。因此,本研究旨在优化血液中DOA I Plus的截止浓度,并比较LC-MS/MS结果。使用Randox Evidence DOA I Plus阵列对680份真实的法医全血样本进行滥用药物筛选,并通过LC-MS/MS进行确认。关于制造商建议的阈值,680份真实血液样本中有139份对一种或多种分析物呈阳性,541份呈阴性。LC-MS/MS确认的139份阳性血液样本中,几乎所有的一种或多种分析物均为真阳性,541份阴性血液样本中有522份为真阴性。总的敏感性和特异性分别为87.8%和99.6%。详细考虑了四氢大麻酚,并进行了受试者-操作员特性曲线分析,以确定四氢大麻醚的最佳截止值,因为根据制造商的建议阈值,它占所有阳性结果的78%。THC的最佳阈值是在浓度为23时确定的 ng/mL,而其他参数的这些值是按照制造商的建议定义的。
{"title":"Optimization of biochip assay for illegal substances on drug abusers' whole blood: Randox Evidence vs LC-MS/MS.","authors":"Pinar Efeoglu Ozseker, Sevinc Puren Yucel, Nebile Daglioglu","doi":"10.1080/15321819.2023.2189451","DOIUrl":"10.1080/15321819.2023.2189451","url":null,"abstract":"<p><p>Immunoassay tests are used in clinical and forensic toxicology laboratories to determine illicit drug use in biological samples. Therefore, this study aims to optimize the cutoff concentrations of DOA I Plus in the blood and compare the LC-MS/MS results. 680 authentic forensic whole blood specimens with Randox Evidence DOA I Plus array were screened for drug of abuse and confirmed by LC-MS/MS. Regarding the manufacturer recommended threshold values, 139 out of 680 authentic blood samples were positive for one or more analytes, while 541 were negative. Nearly all of the 139 positive blood samples confirmed by LC-MS/MS were true positive for one or more analytes, while 522 of the 541 negative blood samples were true negative. The overall sensitivity and the specificity were 87.8% and 99.6%, respectively. THC was considered in detail, and a receiver operator characteristic curve analysis was performed to determine the optimum cutoff for THC, as it accounts for 78% of all positive results according to the manufacturer's recommended thresholds. The optimal threshold value for THC was determined at a concentration of 23 ng/mL, while these values for other parameters were defined as recommended by the manufacturer.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"44 4","pages":"313-325"},"PeriodicalIF":0.0,"publicationDate":"2023-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9842012","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 : 2023-07-04DOI: 10.1080/15321819.2023.2224865
Asmaa El-Shershaby, Nahla Hussein, Esraa Ali, Amr El-Hakim, Ashraf Tabll, Mohamed Shaheen, Ibrahim Ali, Mahmoud Elshall, Yasser Shahein
Several diagnostic measures have been employed to precisely detect the SARS-CoV-2 viral infection using viral antigens, nucleic acids, and other serological approaches. The sensitivity and specificity of the serological tests remain a challenging need. Here, we describe the detection of human anti-SARS-CoV-2 IgG and IgM antibodies qualitatively through two optimized in-house ELISA and lateral flow immunoassay. Both approaches are based on the prokaryotic expression of 50 kDa SARS-CoV-2 recombinant nucleocapsid protein. This SARS-CoV-2rN-6×His was used either to coat ELISA plates or to be conjugated to gold nanoparticles followed by colorimetric detection of bound human IgG or IgM. In the LFA, we show the optimization of nanoparticle size, protein-binding capacity, membrane treatment, and finally testing the potential capacity of using either the optimized ELISA or LFA in detecting antibodies raised against viral infection. Assessment of both methods was carried out using human sera-positive and negative SARS-CoV-2 antibodies. The ELISA and LFA tests showed 86%, 96.5% sensitivity, 92%, 93.75% specificity, 97%, 98.2% PPV, and 64%, 88.2% NPV, respectively. In conclusion, both approaches were able to successfully detect human antibodies against SARS-CoV-2 nucleocapsid protein. The importance of both protocols cannot be overstated in the detection and diagnosis of viral infections, especially in developing countries.
{"title":"Comparative assessment of homemade ELISA and lateral flow assay (LFA)in the rapid, specific and sensitive detection of SARS-CoV-2 anti-nucleocapsid protein in sera of Egyptian patients.","authors":"Asmaa El-Shershaby, Nahla Hussein, Esraa Ali, Amr El-Hakim, Ashraf Tabll, Mohamed Shaheen, Ibrahim Ali, Mahmoud Elshall, Yasser Shahein","doi":"10.1080/15321819.2023.2224865","DOIUrl":"https://doi.org/10.1080/15321819.2023.2224865","url":null,"abstract":"<p><p>Several diagnostic measures have been employed to precisely detect the SARS-CoV-2 viral infection using viral antigens, nucleic acids, and other serological approaches. The sensitivity and specificity of the serological tests remain a challenging need. Here, we describe the detection of human anti-SARS-CoV-2 IgG and IgM antibodies qualitatively through two optimized in-house ELISA and lateral flow immunoassay. Both approaches are based on the prokaryotic expression of 50 kDa SARS-CoV-2 recombinant nucleocapsid protein. This SARS-CoV-2rN-6×His was used either to coat ELISA plates or to be conjugated to gold nanoparticles followed by colorimetric detection of bound human IgG or IgM. In the LFA, we show the optimization of nanoparticle size, protein-binding capacity, membrane treatment, and finally testing the potential capacity of using either the optimized ELISA or LFA in detecting antibodies raised against viral infection. Assessment of both methods was carried out using human sera-positive and negative SARS-CoV-2 antibodies. The ELISA and LFA tests showed 86%, 96.5% sensitivity, 92%, 93.75% specificity, 97%, 98.2% PPV, and 64%, 88.2% NPV, respectively. In conclusion, both approaches were able to successfully detect human antibodies against SARS-CoV-2 nucleocapsid protein. The importance of both protocols cannot be overstated in the detection and diagnosis of viral infections, especially in developing countries.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"44 4","pages":"338-359"},"PeriodicalIF":0.0,"publicationDate":"2023-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10213509","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 : 2023-05-04DOI: 10.1080/15321819.2022.2138718
Mohammad Mohammadi, Mohammad Zangooei, Ebrahim Abbasi, Saeed Ebrahimi Fana, Mahdi Aminian
Tetanus is an acute and often fatal infectious disease caused by Clostridium tetani. Tetanus toxin (TT) is responsible for spastic paralysis observed in tetanus. Anti-tetanus antibodies obtained from horses and humans are the most antitoxins used for tetanus treatment, although some clinical side effects and disadvantages have been reported in their application. The aim of this study is the production of anti-TT IgY and evaluation of its protective effects in a mouse model. Anti-TT IgY was purified from the egg yolk using PEG6000 precipitation and water dilution methods, and its purity was verified by SDS-PAGE. Finally, the potency of purified anti-TT IgY in neutralizing the lethal effects of TT was studied in vivo using a mouse model. PEG6000 precipitation method had better results. Animal studies showed that the purified IgY neutralized the toxic effects of 100 MLD of TT and multiple intravenous-dose injections of anti-TT IgY also had a continuous effect of TT neutralization. The purified anti-TT IgY was effective in neutralizing the lethal activity of TT in a mouse model. Our results suggested that IgY could be an alternative therapeutic source for the management of tetanus in the future.Abbreviations Anti-TT, Anti-tetanus toxin; ELISA, Enzyme-linked immunosorbent assay; IgY, Immunoglobulin Y; MLD, Minimum lethal dose; PBS, Phosphate buffer solution; PEG, Polyethylene glycol; SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis; TIG, Tetanus immune globulin; TT, Tetanus toxin; WD, Water dilution; RT, Room temperature.
{"title":"Production of anti-tetanus toxin IgY and study of its protective effects in a mouse model.","authors":"Mohammad Mohammadi, Mohammad Zangooei, Ebrahim Abbasi, Saeed Ebrahimi Fana, Mahdi Aminian","doi":"10.1080/15321819.2022.2138718","DOIUrl":"https://doi.org/10.1080/15321819.2022.2138718","url":null,"abstract":"<p><p>Tetanus is an acute and often fatal infectious disease caused by <i>Clostridium tetani</i>. Tetanus toxin (TT) is responsible for spastic paralysis observed in tetanus. Anti-tetanus antibodies obtained from horses and humans are the most antitoxins used for tetanus treatment, although some clinical side effects and disadvantages have been reported in their application. The aim of this study is the production of anti-TT IgY and evaluation of its protective effects in a mouse model. Anti-TT IgY was purified from the egg yolk using PEG6000 precipitation and water dilution methods, and its purity was verified by SDS-PAGE. Finally, the potency of purified anti-TT IgY in neutralizing the lethal effects of TT was studied <i>in vivo</i> using a mouse model. PEG6000 precipitation method had better results. Animal studies showed that the purified IgY neutralized the toxic effects of 100 MLD of TT and multiple intravenous-dose injections of anti-TT IgY also had a continuous effect of TT neutralization. The purified anti-TT IgY was effective in neutralizing the lethal activity of TT in a mouse model. Our results suggested that IgY could be an alternative therapeutic source for the management of tetanus in the future.<b>Abbreviations</b> Anti-TT, Anti-tetanus toxin; ELISA, Enzyme-linked immunosorbent assay; IgY, Immunoglobulin Y; MLD, Minimum lethal dose; PBS, Phosphate buffer solution; PEG, Polyethylene glycol; SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis; TIG, Tetanus immune globulin; TT, Tetanus toxin; WD, Water dilution; RT, Room temperature.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"44 3","pages":"283-295"},"PeriodicalIF":0.0,"publicationDate":"2023-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9259788","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}