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Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis
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作者 Dawala Koromtili Oumar Matthew Mutinda Munyao +15 位作者 Anne Wanjiru Mwangi Rebecca Wanjiku Waihenya Peter Kipkemboi Rotich Robinson Mugasiali Irekwa Tonny Teya Nyandwaro Caroline Wangui Njoroge Joanne Jepkemei Yego Primrose Muthoni Ndungu Sharlene Kerubo Mageto Damaris Mutethya Kilei Otilmoi Poul Stephen Nicole Sian Tanchu Grace Ngendo Kanyita shingo inoue Lucy Gitau Samson Muuo Nzou 《American Journal of Molecular Biology》 CAS 2023年第2期127-139,共13页
Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of... Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of the primary phase are not specific, the existing diagnostic methods have low sensitivity and specificity, and the available drugs have some toxicity. New, robust, and cost-effective techniques are needed for the early identification of parasites. This study aimed to assess the sensitivity and specificity of two different types of polyclonal antibodies against T. b. gambiense using antigen detection ELISA. Polyclonal antibodies against the expressed proteins Tbg I2 and Tbg I17 were produced using New Zealand white rabbits. The antibody titer measured was greater than 32 g/L after the 3<sup>rd</sup> immunization for the expressed protein Tbg I2. For the expressed protein Tbg I17, the antibody titer measured was greater than 32 g/L after the 4<sup>th</sup> immunization. The sensitivity and specificity of the Tbg I2 polyclonal antibody confirmed with Polymerase Chain Reaction (PCR) as gold standard were respectively 89.5% and 80.6%, while for the Tbg I17 polyclonal antibody, the sensitivity and specificity were respectively 92.1% and 88.9%. The area under the curve for the Tbg I2 polyclonal antibody was 0.90 ± 0.032, while for the Tbg I17 polyclonal antibody, the area under the curve was 0.92 ± 0.0. The Tbg I17 polyclonal antibody produced in New Zealand white rabbits has good sensitivity and good specificity;it can be successfully used in the diagnosis of HAT. 展开更多
关键词 Human African Trypanosomiasis Polyclonal Antibody Tbg I2 Expressed Protein Tbg I17 Expressed Protein Sensitivity SPECIFICITY
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miR-122-5p as a novel biomarker for alpha-fetoproteinproducing gastric cancer 被引量:9
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作者 Suguru Maruyama Shinji Furuya +10 位作者 Kensuke Shiraishi Hiroki Shimizu Hidenori Akaike Naohiro Hosomura Yoshihiko Kawaguchi Hidetake Amemiya Hiromichi Kawaida Makoto Sudo shingo inoue Hiroshi Kono Daisuke Ichikawa 《World Journal of Gastrointestinal Oncology》 SCIE CAS 2018年第10期344-350,共7页
AIM To investigate the clinical utility of alpha-fetoprotein(AFP)-producing gastric cancer(AFPGC)-specific microRNA(mi RNA)for monitoring and prognostic prediction of patients.METHODS We performed a comprehensive miRN... AIM To investigate the clinical utility of alpha-fetoprotein(AFP)-producing gastric cancer(AFPGC)-specific microRNA(mi RNA)for monitoring and prognostic prediction of patients.METHODS We performed a comprehensive miRNA array-based approach to compare miRNA expression levels between AFP-positive and AFP-negative cells in three patients with primary AFPGC.We next examined the expression levels of the selected miRNAs in five AFPGC and ten non-AFPGC tissue samples by quantitative reverse transcription-polymerase chain reaction to validate their utility.We also investigated the expression levels of the selected miRNA not only in tissue but also in plasma samples.Moreover,we investigated the relationship between plasma AFP levels and plasma selected miRNA expression levels,and also investigated the correlation of the selected miRNA expression levels and malignant potential.RESULTS Among the five miRNAs selected from the miRNA array results,the expression levels of miR-122-5p were significantly higher in the AFPGC patients than in the non-AFPGC patients(P<0.05).In tissue samples,mi R-122-5p expression level tended to be lower in the non-AFPGC tissue than the normal gastric mucosa.Conversely,in the AFPGC tissue,miR-122-5p expression level was significantly higher in the AFPGC tissue than both the normal gastric mucosa and the nonAFPGC tissue samples(P<0.05).Plasma mi R-122-5p expression levels were also significantly higher in the AFPGC patients than the health volunteers and the nonAFPGC patients(P<0.05)and were strongly correlated with plasma AFP levels(r=0.7975,P<0.0001).Moreover,the correlation of miR-122-5p expression in tissue samples with malignant potential was stronger than that of plasma AFP level in the AFPGC patients.In contrast,no correlation was found between mi R-122-5p expression levels and liver metastasis in the non-AFPGC patients.CONCLUSION miR-122-5p might be a useful biomarker for early detection and disease monitoring in AFPGC. 展开更多
关键词 GASTRIC CANCER ALPHA-FETOPROTEIN Alphafetoprotein producing GASTRIC CANCER MicroRNA miR-122-5p
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日本脑炎ML-17两个变异株的基因组序列及突变分析(英文)
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作者 金玉明 Afjal Hossain Khan +3 位作者 Mohammed Alimul Islam Takeshi Nabeshima shingo inoue Kouichi Morita 《中国热带医学》 CAS 2006年第8期1340-1344,共5页
目的鉴别日本脑炎疫苗株ML-17的两个大宽变异株(ML-17L2和ML-17L4)的存在,并通过全基因组序列测定分析引起斑形和毒力改变的基因突变。方法首先对疫苗株ML-17在传代过程中出现的多斑ML-17sh的两个大宽变异株进行纯化和鉴定,再对两... 目的鉴别日本脑炎疫苗株ML-17的两个大宽变异株(ML-17L2和ML-17L4)的存在,并通过全基因组序列测定分析引起斑形和毒力改变的基因突变。方法首先对疫苗株ML-17在传代过程中出现的多斑ML-17sh的两个大宽变异株进行纯化和鉴定,再对两个大斑变异株进行全基因组序列测定和全编码氨基酸序列推导。并与小斑疫苗株ML-17及其大斑亲代野毒株JaOH0566的全核苷酸序列和全编码氨基酸序列进行比较,分析引起宽形改变的基因突变。并应用小白鼠试验检查ML-17变异前后的神经侵袭力和毒力变化。结果全基因组序列分析显示两个大斑变异株的核苷酸总数均为10977个,与ML-17和JaOH0566相比较,分剐有33和34个核苷酸变异散在分布于ML-17L2和ML-17L4的全基因组中,其中又分别有12和13个核苷酸导致了氨基酸的改变。在ML-17L2和ML-17L4的衣壳蛋白区没有氨基酸变异,但在包膜蛋白区各有一个氨基酸变异;在5’端非编码区没有核苷酸变异,但在3’端非编码区有3~4个核苷酸变异,且各增加了一个额外的10699核苷酸。ML-17和ML-17sh的小鼠LD50分别为大于1×10^6FFU和481FFU。变异后毒力明显增强。结论发生在两个大斑变异株的第5非结构蛋白区和其它区的氨基酸返祖变异(变回与JaOH0566相同)和新变异,可能引起了宽彤的改变;而全部5个氨基酸返祖变异和2—3个氨基酸新变异可能与其毒力改变有关。本次研究为ML-17L2和ML-17L4的斑形和毒力改变提供了分子遗传学基础。 展开更多
关键词 日本脑炎 ML-17减毒活疫苗株 全核苷酸序列测定 突变
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