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Rapid Detection of rpoB Gene Mutations in Rif-resistant M.tuberculosis Isolates by Oligonucleotide Microarray 认领 引用 被引量：9
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作者 AI-HUA SUN XING-LI FAN +3 位作者 LI-WEI LI LI-FANG WANG WEN-YING AN JIE YAN 《Biomedical and Environmental Sciences》 SCIE CAS 2009年第3期253-258,共6页
Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DN... Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates. 展开更多
关键词 Mycobacterium tuberculosis Rifampin resistance rpoB gene / site mutation Oligonucleotide microarray/detection
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Development and application of antibody microarray for white spot syndrome virus detection in shrimp 认领 引用 被引量：3
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作者 徐晓丽 绳秀珍 战文斌 《Chinese Journal of Oceanology and Limnology》 CSCD 2011年第5期930-941,共12页
Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the re... Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the requirements. In this study, we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples. First, seven slides each with different modifications were characterized by atomic force microscope, and were compared in the efficiency of immobilizing proteins. Of the seven, 3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size. A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides, and then the mieroarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody. The results were measured by a laser chipscanner and analyzed with software. To obtain satisfied fluorescence signal intensity, optimal conditions were searched. The detection limit of the antibody microarray for WSSV is 0.62μg/mL, with a woven long shelf life for 6 months at 4℃ or 8 months at -20℃. Furthermore, concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection. These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV. 展开更多
关键词 shrimp white spot syndrome virus antibody microarray agarose gel detection
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Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters 认领 引用 被引量：2
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作者 陈先锋 周前进 +6 位作者 段维军 周成旭 段丽君 张慧丽 孙爱丽 严小军 陈炯 《Chinese Journal of Oceanology and Limnology》 CSCD 2016年第1期86-101,共16页
Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully develop... Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples. 展开更多
关键词 ballast waters DNA microarray harmful algae limit of detection multiplex PCR seaport waters
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Detection of microarray protein biomolecules by oblique-incidence reflectivity difference technique without labelling agents 认领 引用 被引量：2
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作者 张洪艳 陆珩 +5 位作者 李巍 梁汝强 金奎娟 周岳亮 阮康成 杨国桢 《Chinese Physics B》 SCIE EI CAS 2008年第6期2288-2291,共4页
This paper reports that the detection to the protein in microarray format is carried out by oblique-incidence reflectivity difference （OI-RD） analysis without any labelling agents. The OI-RD intensities not only dep... This paper reports that the detection to the protein in microarray format is carried out by oblique-incidence reflectivity difference （OI-RD） analysis without any labelling agents. The OI-RD intensities not only depend on the protein structure, but also vary with the protein concentration. The results indicate that this method should have potential application in detection of biochemical processes. The high throughout and in situ detection can be achieved by this method with further improving of the experimental system. 展开更多
关键词 detection OI-RD protein microarray
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Identification of SRSF2 as a potential immunodiagnostic biomarker for osteosarcoma:A study based on focused protein microarray 认领 引用
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作者 Yan Ma Lei Wan +7 位作者 Yong-Yong Zhang Ya-Ge Luo Ning Yang Fei-Fei Pu Malik Ihsan Ullah Khan Man Liu Jin-Shan Liu Ji-Tian Li 《World Journal of Clinical Oncology》 2026年第5期153-164,共12页
BACKGROUND Osteosarcoma(OS)is a primary solid tumor of bone.Due to the insidious onset and malignancy of OS,it is necessary to diagnose OS earlier.AIM To search for an optimal diagnostic biomarker,such as autoantibody... BACKGROUND Osteosarcoma(OS)is a primary solid tumor of bone.Due to the insidious onset and malignancy of OS,it is necessary to diagnose OS earlier.AIM To search for an optimal diagnostic biomarker,such as autoantibody against tumor associated antigen(TAA),for early diagnosis of OS.METHODS Differential expression of biomarkers in sera from 28 patients with OS and 49 normal human sera were screened by a focused protein microarray with 154 human recombinant proteins.Enzyme-linked immunosorbent assay(ELISA)was applied to validate the three potential biomarkers from 49 OS patients matched with 49 normal controls and 39 osteochondroma.The diagnostic value of each TAA for OS was analyzed by receiver operating characteristic curve.Western blotting and immunohistochemistry were used to verify the results of ELISA.RESULTS Based on the protein microarray,three differentially expressed TAAs[G-protein subunit alpha 11,serine/arginine-rich splicing factor 2(SRSF2),and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha]were screened out,whose content was higher than in normal human sera.According to the results of ELISA,anti-SRSF2 autoantibody showed higher expression in OS than in normal controls,with sensitivity,specificity,and area under the curve of 16.33%,95.92%,and 0.648,respectively.The ELISA results were confirmed by western blotting and immunohistochemistry.CONCLUSION Autoantibody is potential serological biomarker in detecting OS.Expression of anti-SRSF2 autoantibody has the potential to distinguish OS from the normal individuals. 展开更多
关键词 Osteosarcoma Tumor associated antigen Protein microarray Detection SRSF2 Biomarker
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Using oligonucleotide suspension arrays for laboratory identification of bacteria responsible for bacteremia 认领 引用 被引量：4
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作者 Xiao-li HOU Han-liang JIANG +3 位作者 Qing-yi CAO Li-ying ZHAO Barbara J. CHANG Zhi CHEN 《Journal of Zhejiang University-SCIENCE B》 SCIE CAS 2008年第4期291-298,共8页
The aim of this study was to develop and validate an oligonucleotide suspension array for rapid identification of 15 bacterial species responsible for bacteremia, particularly prevalent in Chinese hospitals. The multi... The aim of this study was to develop and validate an oligonucleotide suspension array for rapid identification of 15 bacterial species responsible for bacteremia, particularly prevalent in Chinese hospitals. The multiplexed array, based on the QIAGEN LiquiChip Workstation, included 15 oligonucleotide probes which were covalently bound to different bead sets. PCR amplicons of a variable region of the bacterial 23S rRNA genes were hybridized to the bead-bound probes. Thirty-eight strains belonging to 15 species were correctly identified on the basis of their corresponding species-specific hybridization profiles. The results show that the suspension array, in a single assay, can differentiate isolates over a wide range of strains and species, and suggest the potential utility of suspension array system to clinical laboratory diagnosis. 展开更多
关键词 Oligonucleotide array Bacteremia 23S rRNA Multiplexed detection
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SELECTION AND THEIR ANTITUMOR ACTIVITY OF ANTISENSE OLIGONUCLEOTIDES TARGETING MESSENGER RNA OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR 2 认领 引用
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作者 郑素军 林汝仙 +4 位作者 夏云 伯晓晨 任红 钟森 王升启 《Chinese Journal of Cancer Research》 CAS 2005年第3期161-170,共10页
Objective： To select the antisense oligonucleotides （asONs） which hybridize with the mRNA of vascular endothelial growth factor receptor2 （VEGFR2, also named as kinase insert domain-containing receptor：KDR） in a... Objective： To select the antisense oligonucleotides （asONs） which hybridize with the mRNA of vascular endothelial growth factor receptor2 （VEGFR2, also named as kinase insert domain-containing receptor：KDR） in an effective and specific way, and to investigate their antitumor activity in MCF-7 cells. Methods： The effective antisense oligonucleotides were chosen by computer prediction combined with oligonucleotide microarrays. The inhibition effect on MCF-7 cells proliferation was measured by MTT; and VEGFR2 expression was surveyed by Western-blotting and RT- PCR. Results： Using predicting secondary structure of VEGFR2 mRNA with RNA folding program, computer prediction designed 30 antisense oligonucleotide probes that were directed to local loose regions of RNA structure. In 30 probes, 4（4/30, 13.33%） antisense oligonucleotides showed strong hybridization intensities in oligonucleotide microarrays test and were selected. All these antisense oligonucleotides targeting 4 different sites of VEGFR2 mRNA lowered the level of VEGFR2 mRNA and protein present in MCF-7 cells. Proliferation of MCF-7 cells was reduced by 4 antisense oligonucleotides, respectively, in which asON1 was the most effective, with the inhibitory rates being 53.06% at 0.8 I.tmol/L. Conclusion： Combination of computer prediction with oligonucleotide microarrays is an effective way in selecting optimal antisense oligonucleotides. The antisense oligonucleotides showed good correlation between their antitumor activity and the hybridization intensities. The antisense oligonucleotides targeting VEGFR2 mRNA demonstrated prominent antitumor role in vitro. 展开更多
关键词 Target selection Computer prediction Oligonucleotide microarrays Antisense oligonucleotide VEGFR2/KDR
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Novel biosensor-based microarray assay for detecting rs8099917 and rs12979860 genotypes 认领 引用 被引量：2
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作者 Pei-Yuan Li Xiao-Jun Zhou +3 位作者 Lan Yao Xin-Hua Fang Jiang-Nan Ren Jia-Wu Song 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第44期6481-6488,共8页
AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 genotypes.METHODS:Four probes specific for rs8099917C/T or rs12979860G/T detection and three sets of quality control... AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 genotypes.METHODS:Four probes specific for rs8099917C/T or rs12979860G/T detection and three sets of quality control probes were designed,constructed and arrayed on an optical biosensor to develop a microarray assay.Two sets of primers were used in a one tube polymerase chain reaction(PCR) system to amplify two target fragments simultaneously.The biosensor microarray contained probes that had been sequenced to confirm that they included the rs8099917C/T or rs12979860G/T alleles of interest and could serve as the specific assay standards.In addition to rehybridization of four probes of known sequence,a total of 40 clinical samples collected from hepatitis C seropositive patients were also tested.The target fragments of all 40 samples were amplified in a 50 μL PCR system.Ten μL of each amplicon was tested by BBM assay,and another 40 μL was used for sequencing.The agreement of the results obtained by the two methods was tested statistically using the kappa coefficient.The sensitivity of the BBM assay was evaluated using serial dilutions of ten clinical blood samples containing 10 3-10 4 white cells/μL.RESULTS:As shown by polyacrylamide gel electrophoresis,two target segments of the interleukin 28Bassociated polymorphisms(SNPs) were successfully amplified in the one-tube PCR system.The lengths of the two amplified fragments were consistent with the known length of the target sequences,137 and 159 bps.After hybridization of the PCR amplicons with the probes located on the BBM array,the signals of each allele of both the rs8099917 SNPs and rs12979860 SNPs were observed simultaneously and were clearly visible by the unaided eye.The signals were distinct from each other,could be interpreted visually,and accurately recorded using an ordinary digital camera.To evaluate the specificity of the assay,both the plasmids and clinical samples were applied to the microarray.First,30 PCR amplicons of the various SNP alleles were hybridized on the BBM microarray.Full agreement between plasmids and the BBM assay was observed,with 30/30 correct matches(100%).The kappa value for the BBM assay with plasmids was 1.00(P 0.75 was defined as substantial agreement).The BBM assay and sequencing had similar specificities for detection and identification of the two SNPs and their alleles.The sensitivity evaluation showed that the BBM assay could detect and identify SNP sequences present in blood samples containing as few as 10 2 white blood cells/μL.CONCLUSION:This biosensor microarray assay was highly specific,sensitive,rapid and easy to perform.It is compatible with clinical practice for detection of rs8099917 and rs12979860. 展开更多
关键词 Biosensor-based microarray Hepatitis C vi-rus rs8099917 rs12979860 Detection Assay
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Detection and Characterization of β-Lactamase Encoding Genes in Carbapenem Non-Susceptible Gram-Negative Bacteria and Susceptibility of Isolates to Ceftazidime-Avibactam at a New York City Community Hospital 认领 引用
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作者 Carl Urban Rita Colon-Urban +5 位作者 Vincent J. LaBombardi Noriel Mariano Nishant Prasad Geeti Dhillon Marina Guralnik Sorana Segal-Maurer 《Open Journal of Medical Microbiology》 2016年第4期150-157,共8页
A surveillance study was undertaken to identify prominent β-lactamase encoding genes in 131 carbapenem non-susceptible gram-negative clinical isolates at a New York City community hospital. KPC carbapenemases were de... A surveillance study was undertaken to identify prominent β-lactamase encoding genes in 131 carbapenem non-susceptible gram-negative clinical isolates at a New York City community hospital. KPC carbapenemases were detected in 89% of Enterobacteriaceae as well as additional TEM, SHV, and CTX-M class A enzymes. OXA-23 and OXA-24 were the prevalent class D carbapenemases identified in Acinetobacter species. One OXA-23 in M. morganii and one OXA-48 in K. pneumoniae were also identified. Among class C β-lactamases CMY, ACT/MIR, DHA, and FOX were detected. The in vitro activity of ceftazidime-avibactam by E-test methodology was tested with minimal inhibitory concentrations (MIC) of ≤3 μg/ml for 97.8% of all Enterobacteriaceae, MIC50/90 of 16/>256 μg/ml for carbapenem non-susceptible Acinetobacter, and 3/6 μg/ml for carbapenem non-susceptible Pseudomonas aeruginosa. Periodic surveillance of isolates to characterize current and emerging β-lactamase genotypes present in local isolates may help identify outbreak situations, provide assistance to infection control and antibiotic stewardship programs, and potentially improve patient outcomes. 展开更多
关键词 Carbapenem Non-Susceptible Check-MDR CT103 XL Microarray β-Lactamase Detection Resistance Mechanisms Ceftazidime-Avibactam
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Genome-wide identification of gene expression in the epididymis of infertile rat induced by alpha-chlorohydrin using oligonucleotide microarray 认领 引用
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作者 XIE Shu-wu ZHU Yan +5 位作者 MA Li LI Zhi-ling GUI You-lun LU Ying-ying ZHAO Zhi-fang CAO Lin 《沈阳药科大学学报》 CAS 北大核心 2008年第S1期111-112,共2页
Objective To establish a rat model of sterility associated with epididymis and epididymal gene expression profiles relation to fertility by alpha-chlorohydrin.Methods Rats were treated with 10 mg·kg-1·d-1.al... Objective To establish a rat model of sterility associated with epididymis and epididymal gene expression profiles relation to fertility by alpha-chlorohydrin.Methods Rats were treated with 10 mg·kg-1·d-1.alpha-chlorohydrin for 10 consecutive days.Sperm maturation and other fertility parameters were analyzed.The sperm motility and morphology were evaluated by computer-assisted sperm analysis(CASA);sperm survival rate was assessed by SYBR-14 and propidium iodide(PI)fluorescent staining;the weights of testes,epididymides,prostates and seminal vesicles were determined by electronic balance;histological examination of above tissues were evaluated by HE staining;and serumal dihydrotestosterone(DHT)and testosterone(T)of rats were detected by enzyme-labeled immunoassay.Each male rat was paired with 2 female rats in proestrus.Female rats were examined the next morning for the presence of sperm in vaginal smears and underwent a cesarean section on day 12 of gestation.Finally the reproductive indices were calculated as follows:copulation index(number of sperm positive femalesumber of pairings),pregnancy index(number of pregnanciesumber of sperm positive females),and fertility index(number of pregnanciesumber of pairings).After that we used Affymetrix Rat 2302.0 oligo-microarray to identify epididymal special genes associated with fertility.Finally,we validated some of these genes by Real-Time quantitative polymerase chain reaction.Results The motility of spermatozoa from the cauda epididymidis of treated rats showed a significant decrease in percentage of motile,progressively motile sperm,and sperm survival rate.At the same time,the morphology of cauda epididymal spermatozoa was also adversely affected by the treatment.In addition,the serumal androgen levels of treated animals weren't changed compared with the control group.Accordingly,matings with treated males resulted in no successful pregnancy.Then,we classified general functions of the down or up regulated epididymal genes by chlorhydrin with the GeneSpring gene ontology(GO)analysis,which are involved in macromolecular metabolism and transport,primary metabolism process,cell metabolism,biological process regulation,immunology regulation,ion combination,hydratase and oxidoreductase activity.Among all the different expressed genes,we analyzed and screened the down-regulated genes associated with glucose,lipid,protein and other energy metabolism,which are considered as the major ACH action targets.Simultaneously,the up-regulated genes by chlorhydrin were detected and their characters of negative regulated sperm maturation and fertility analyzed,which are apoptosis and immune-related genes and not reported before.Conclusions We established male infertile rat model with ACH(10 mg·kg-1·d-1,po,10 days)through evaluating changes of sperm motility and morphology,mating index,fertility index and pregnancy index.Simultaneously,the ACH didn't affect the major androgen(T and DHT)metabolism and sexual ability,which is considered as the best way for male contraception.Then we determined the down-regulated epididymal genes relation to substance metabolism,which can affect the epididymal sperm maturation and presumed the major antifertility targets by ACH.Further more,we found and analyzed the epididymal up-regulated genes associated with apoptosis and immune functions,which maybe the new possible sites of action by ACH and other male antifertility agents. 展开更多
关键词 male infertile rat ACH antifertility agents oligonucleotide microarray
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Design and application of 60mer oligonucleotide microarray in SARS coronavirus detection 认领 引用 被引量：4
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作者 SHI Rong MA Wenli +6 位作者 WU Qinghua ZHANG Bao SONG Yanbin GUO Qiuye XIAO Weiwei WANG Yan ZHENG Wenling 《Chinese Science Bulletin》 2003年第12期1165-1169,共5页
The 60mer oligonucleotide microarray was designed and applied to detecting of SARS (severe acute res-piratory syndrome) coronavirus. Thirty 60mer specific oligos were designed to cover the whole genome of the first su... The 60mer oligonucleotide microarray was designed and applied to detecting of SARS (severe acute res-piratory syndrome) coronavirus. Thirty 60mer specific oligos were designed to cover the whole genome of the first submit-ted coronavirus strain, according to the sequence of TOR2 (GENEBANK Accession: AY274119). These primers were synthesized and printed into a microarray with 12×12 spots. RNAs were extracted from the throat swab and gargling fluid of SARS patients and reverse-transcripted into the double strand cDNAs. The cDNAs were prepared as re-stricted cDNA fragments by the restriction display (RD) technique and labeled by PCR with the Cy5-universal primer. The labeled samples were then applied to the oligo microar-ray for hybridization. The diagnostic capability of the mi-croarray was evaluated after the washing and scanning steps. The scanning result showed that samples of SARS patients were hybridized with multiple SARS probes on the microar-ray, and there is no signal on the negative and blank controls. These results indicate that the genome of SARS coronavirus can be detected in parallel by the 60mer oligonucleotide mi-croarray, which can improve the positive ratio of the diagno-sis. The oligo microarray can also be used for monitoring the behavior of the virus genes in different stages of the disease status. 展开更多
关键词 SARS coronavirus oligonucleotide microarray RD tech-nique fluorescent labeling molecular hybridization.
流式分析技术快速精准检测酸奶中嗜热链球菌活菌 认领 引用 被引量：1
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作者 蔡小雨 代森迪 +6 位作者 丁文龙 王梓权 王蒙 齐聪岩 张伟 刘思渊 隋志伟 《食品科学》 EI CAS CSCD 北大核心 2026年第7期109-118,共10页
为实现对嗜热链球菌活菌数的高效精准定量,本研究创新性地将自主设计的寡核苷酸探针(novel oligonucleotide probe,NOP)与叠氮溴化丙锭(propidium monoazide,PMA)核酸染料相结合,利用流式细胞术(flow cytometry,FCM)构建PMA-NOP-FCM检... 为实现对嗜热链球菌活菌数的高效精准定量,本研究创新性地将自主设计的寡核苷酸探针(novel oligonucleotide probe,NOP)与叠氮溴化丙锭(propidium monoazide,PMA)核酸染料相结合,利用流式细胞术(flow cytometry,FCM)构建PMA-NOP-FCM检测方法。其中PMA在光敏条件下与死菌DNA不可逆共价交联,使死菌携带红色荧光,嗜热链球菌与NOP特异性杂交携带绿色荧光,最后通过流式分析技术识别计数嗜热链球活菌。该方法特异性良好,能够准确识别并定量嗜热链球菌活菌;方法的定量限为8.3×104 cells/g,在104~108 cells/g与平板计数法具有良好的线性关系;相对标准偏差为4.68%,具有较高精密度;检测时间为2 h,显著优于平板计数法。该方法在酸奶加标样品中的回收率为91.61%~106.17%,经验证可有效用于不同市售品牌酸奶的检测。PMA-NOP-FCM方法为酸奶工业化生产中菌群活性监控提供了高精度、高效率的技术解决方案,对酸奶质量控制具有重要实践意义。 展开更多
关键词 流式细胞术 嗜热链球菌 寡核苷酸探针 活菌检测 酸奶
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Label-free detection of hybridization of oligonucleotides by oblique-incidence reflectivity difference method 认领 引用 被引量：8
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作者 YUAN Kun WANG Xu +7 位作者 LU Heng WEN Juan LU HuiBin ZHOU YueLiang JIN Kui-Juan YANG GuoZhen LI Wei RUAN KangCheng 《Science China(Physics,Mechanics & Astronomy)》 SCIE EI CAS 2010年第8期1434-1437,共4页
The microarrays of 20-base oligonucleotide with different concentrations are detected before and after hybridization by the oblique-incidence reflectivity difference (OI-RD) method. The experimental results prove that... The microarrays of 20-base oligonucleotide with different concentrations are detected before and after hybridization by the oblique-incidence reflectivity difference (OI-RD) method. The experimental results prove that OI-RD is a label-free method which can not only distinguish the concentration difference of oligonucleotides before and after the hybridization but also detect the hybridization of short oligonucleotides. At present the OI-RD method can detect 0.39 μmol/L 20-base oligonucleotide or less. These results suggest that the OI-RD method is a promising and potential technique for label-free detection of biological microarrays. 展开更多
关键词 oblique-incidence reflectivity difference microarray detection label-free detection oligonucleotides microarray
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用于生物标志物微阵列芯片检测的高灵敏度成像系统 认领 引用
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作者 孙飞跃 郑璐璐 《光学仪器》 CAS 2026年第3期34-40,49,共7页
本文研究了一种用于生物标志物微阵列芯片检测的高灵敏度成像系统。该系统以共聚焦型光路为成像基础,结合维纳滤波反卷积算法,以及阈值背景矫正处理,有效降低了点扩散函数引起的噪声及随机噪声。该系统对阿尔茨海默病的3种生物标志物微... 本文研究了一种用于生物标志物微阵列芯片检测的高灵敏度成像系统。该系统以共聚焦型光路为成像基础,结合维纳滤波反卷积算法,以及阈值背景矫正处理,有效降低了点扩散函数引起的噪声及随机噪声。该系统对阿尔茨海默病的3种生物标志物微阵列芯片的检测结果表明:其对Aβ42(β淀粉样蛋白42)、P-tau181(磷酸化tau 181蛋白)、T-tau(总tau蛋白)的检出限分别为8.60、20.60、13.12 pg/mL,各检测曲线的决定系数R2均大于0.99,测量重复性指标小于3.8%。与博奥生物公司的共聚焦芯片扫描仪的对比测试结果显示,该系统的检测限达到了商用共聚焦芯片扫描仪的同等数量级水平,并解决了激光共聚焦扫描成像技术的高复杂性与传统CCD成像灵敏度较低的问题,在疾病现场实时检测与预防等领域中具有广阔的应用空间。 展开更多
关键词 微阵列技术 蛋白质芯片 反卷积 标志物检测
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胎儿先天性心脏病产前基因诊断的研究进展 认领 引用
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作者 金佳 关艳梅 陈静 《中国优生与遗传杂志》 2026年第5期1074-1078,共5页
先天性心脏病(CHD)位于围生期出生缺陷发生率的首位,并有较高的死亡率。其病因复杂,发病机制不清,遗传和环境因素在疾病发展中起着重要作用。合并其他系统畸形的CHD检出染色体异常的概率更高,这类CHD严重影响新生儿的健康,临床预后较差... 先天性心脏病(CHD)位于围生期出生缺陷发生率的首位,并有较高的死亡率。其病因复杂,发病机制不清,遗传和环境因素在疾病发展中起着重要作用。合并其他系统畸形的CHD检出染色体异常的概率更高,这类CHD严重影响新生儿的健康,临床预后较差,给患儿家庭带来了沉重的心理和经济负担,尽早筛查出伴染色体异常的CHD,有助于孕期给予咨询及进一步指导。因此对CHD胎儿进行产前基因诊断及遗传咨询至关重要。随着产前诊断技术的不断发展,胎儿CHD的产前基因诊断取得了显著进展。本文旨在梳理胎儿CHD产前基因诊断的技术发展及临床应用现状,为临床应用提供参考。 展开更多
关键词 先天性心脏病 产前基因诊断 染色体微阵列分析 基因组拷贝数变异检测 全外显子组测序
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Detection of hybridization of protein microarrays using an oblique-incidence reflectivity difference method 认领 引用 被引量：8
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作者 LU Heng WEN Juan +7 位作者 WANG Xu YUAN Kun LU HuiBin ZHOU YueLiang JIN Kui-Juan YANG GuoZhen LI Wei RUAN KangCheng 《Science China(Physics,Mechanics & Astronomy)》 SCIE EI CAS 2010年第7期1230-1233,共4页
Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence refl... Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) method.Two detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained simultaneously.The detection results of hybridization by OI-RD were in accord with that of traditional fluorescent scans.In particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic coordinates.The detection sensitivity of OI-RD can reach 14 fg.These experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis. 展开更多
关键词 protein microarrays oblique-incidence reflectivity difference(OI-RD) label-free detection
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Detection of protein microarrays by oblique-incidence reflectivity difference technique 认领 引用 被引量：4
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作者 WEN Juan LU Heng +7 位作者 WANG Xu YUAN Kun LvHuiBin ZHOU YueLiang JIN KuiJuan YANG GuoZhen LI Wei RUAN KangCheng 《Science China(Physics,Mechanics & Astronomy)》 SCIE EI CAS 2010年第2期306-309,共4页
Biological microarrays with different proteins and different protein concentrations are detected without external labeling by an oblique-incidence reflectivity difference(OIRD)technique.The initial experiment results ... Biological microarrays with different proteins and different protein concentrations are detected without external labeling by an oblique-incidence reflectivity difference(OIRD)technique.The initial experiment results reveal that the intensities of OIRD signals can distinguish the different proteins and concentrations of protein.The OIRD technique promises feasible applications to life sciences for label-free and high-throughput detection. 展开更多
关键词 oblique-incidence reflectivity difference protein microarray label-free detection
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Rhodamine B doped silica nanoparticle labels for protein microarray detection 认领 引用 被引量：3
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作者 WANG Ying LI ZhongHui +3 位作者 ZHONG WenYing LI Hui XU DanKe CHEN HongYuan 《Science China Chemistry》 SCIE EI CAS 2010年第4期747-751,共5页
A core-shell Rhodamine B-doped SiO2 nanoparticle was synthesized and its fluorescent intensity was found to be 1000 times higher than that of individual Rhodamine B molecule.The doped nanoparticles were further conjug... A core-shell Rhodamine B-doped SiO2 nanoparticle was synthesized and its fluorescent intensity was found to be 1000 times higher than that of individual Rhodamine B molecule.The doped nanoparticles were further conjugated with streptavidin and the resulting nanoparticles were used in the detection of reverse-phase protein microarrays,in which human IgG of various concentrations was first immobilized on aldehyde-modified glass slides and then biotinlyated goat anti human IgG as well as the labeled nanoparticles were sequentially conjugated.The calibration curve is linear over the range from 800 fg to 500 pg and the limit of detection is 100 fg,which is 8 times lower than that of streptavidin-labeled Cy3 fluorescent dyes.The dyedoped SiO2 nanoparticles show potentials for the protein array detection. 展开更多
关键词 Rhodamine B SiO2nanoparticles protein detection protein microarrays
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分析应用芯片检测高糖和葡萄糖胺诱导的基因表达对肾系膜细胞氧化应激的预警意义 认领 引用 被引量：1
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作者 余洁玲 梁洁如 钟明芝 《中国实用医药》 2025年第2期76-79,共4页
目的探讨应用芯片检测高糖和葡萄糖胺诱导的基因表达评估肾系膜细胞氧化应激的预警意义和价值。方法选择120例糖尿病肾病(DN)患者为观察组,另选同时间段120例健康体检者为对照组。采用芯片检测高糖和葡萄糖胺诱导的基因表达,检测两组细... 目的探讨应用芯片检测高糖和葡萄糖胺诱导的基因表达评估肾系膜细胞氧化应激的预警意义和价值。方法选择120例糖尿病肾病(DN)患者为观察组,另选同时间段120例健康体检者为对照组。采用芯片检测高糖和葡萄糖胺诱导的基因表达,检测两组细胞氧化应激及肾功能指标。比较两组谷氨酰胺果糖6-磷酸氨基转移酶(GFAT)、细胞外基质(ECM)、转化生长因子α、泛素样结构域蛋白成员1、硫氧还蛋白(TRX)、谷胱甘肽过氧化酶1、代谢型谷氨酸受体1、70 kDa热休克蛋白5基因表达水平,细胞氧化应激[还原型谷胱甘肽(GSH)与氧化型谷胱甘肽(GSSG)的比值(GSH/GSSG)、活性氧自由基]及肾功能指标[尿微量白蛋白、肌酐(Cr)、视黄醇结合蛋白(RBP)、β2微球蛋白(β2-MG)、胱抑素C(Cys C)、尿素氮(BUN)];采用Pearson线性相关分析高糖和葡萄糖胺诱导的基因表达与肾系膜细胞氧化应激的相关性。结果观察组GFAT、ECM、转化生长因子α、泛素样结构域蛋白成员1、TRX、谷胱甘肽过氧化酶1、代谢型谷氨酸受体1、70 kDa热休克蛋白5基因表达水平分别为(1.57±0.32)、(1.61±0.35)、(1.43±0.29)、(1.44±0.32)、(1.63±0.37)、(1.32±0.24)、(1.54±0.27)、(1.58±0.33),高于对照组的(0.63±0.11)、(0.75±0.17)、(0.46±0.07)、(0.78±0.19)、(0.54±0.12)、(0.68±0.14)、(0.43±0.09)、(0.84±0.21)(P<0.05)。观察组GSH/GSSG(6.67±0.11)低于对照组的(9.02±0.07),活性氧自由基(2853.73±62.16)μmol/L、尿微量白蛋白(42.64±3.26)mg/24 h、Cr(213.85±89.41)μmol/L、RBP(114.29±16.25)mg/L、β2-MG(3.05±0.21)mg/L、Cys C(5.31±0.61)mg/L、BUN(12.13±2.14)mmol/L高于对照组的(2325.91±55.79)μmol/L、(19.21±1.52)mg/24 h、(91.19±14.29)μmol/L、(40.24±8.51)mg/L、(1.32±0.06)mg/L、(1.03±0.23)mg/L、(4.39±0.84)mmol/L(P<0.05)。Pearson线性相关分析结果显示:DN患者GFAT、ECM、转化生长因子α、泛素样结构域蛋白成员1、TRX、谷胱甘肽过氧化酶1、代谢型谷氨酸受体1、70 kDa热休克蛋白5基因与活性氧自由基呈正相关,与GSH/GSSG呈负相关(P<0.05)。结论应用芯片能检测高糖和葡萄糖胺诱导的基因表达水平,且与肾系膜细胞氧化应激存在强相关性,能发挥良好的预警作用,为后续诊疗提供依据。 展开更多
关键词 芯片检测 高糖 葡萄糖胺 基因表达 肾系膜细胞 氧化应激 预警意义
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Construction, detection and microarray analysis on the Shigella flexneri 2a sitC mutant 认领 引用 被引量：1
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作者 LIU Moqing LIU Hong +4 位作者 SUN Lilian DONG Jie XUE Ying CHEN Shuxia JIN Qi 《Science China(Life Sciences)》 EI CAS 2005年第3期228-240,共13页
In order to overcome the defects of difficult gene operations in low-copy suicide plasmid pCVD442,Gateway technology was applied in the construction process of recombinant plasmid for gene knockout in this study.With ... In order to overcome the defects of difficult gene operations in low-copy suicide plasmid pCVD442,Gateway technology was applied in the construction process of recombinant plasmid for gene knockout in this study.With this improved knockout system,we inactivated sitC gene,which is associated with iron transport in Shigella flexneri 2a strain 301,to yield the mutant,MTS.The functional detection of the mutant was performed at the level of culture medium,cell and animal experiment,respectively.The gene expression profiles were compared with DNA microarray between the mutant and the wild type under iron-restricted conditions.The results showed that MTS grew obviously less well than the wild-type strains in L broth containing 150μmol/L iron chelator DIP(2,2′-dipyridyl).Addition of iron or manganese to the cultures stimulated the growth of MTS to wild-type levels in rich culture medium.In either the experiment on the ability of intracellular multiplication and cell-to-cell spread in HeLa and U937 cell lines,or the experiment on keratoconjunctivitis in guinea pigs,MTS showed no obvious changes in virulence compared with the parental strain Sf301.When 65μmol/L DIP was added to the cultured HeLa cells,the ability of intracellular multiplication of MTS reduced about 51.6%as compared with that of Sf301.The analysis of expression profiles under iron-limited condition showed that MTS was more sensitive for the change of iron deficiency than Sf301.There are 106 more up-regulated genes in MTS than in wild-type strains,which are involved in membrane transportation,amino acid metabolism and uncategorized function genes,while down-regulated genes are mainly in-volved in energy and carbohydrate metabolism.Under low iron conditions,the expression levels of known iron-transport associated genes generally increased.Additionally,the number of these genes and their increase amplitude in MTS are more than those in Sf301.Together,these results confirmed that Sit iron-transport system is important for the growth of Shigella. 展开更多
关键词 Shigella flexneri 2a gene knockout functional detection microarray analysis.
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