2.Development of a high-throughput suspension microarray technology for detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-beta-estradiol.
Nan LIU ; Pu SU ; Zhi-Xian GAO ; Mao-Xiang ZHU ; Zhi-Hua YANG ; Xiu-Ji PAN ; Fu-Huan CHAO
Chinese Journal of Preventive Medicine 2009;43(6):482-488
OBJECTIVETo establish a novel suspension microarray technology for the detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-beta-estradiol (CAP, CL and E2).
METHODSThe three conjugates that veterinary drug coupled with bovine serum albumin (BSA) were synthesized and identified by ultraviolet (UV) spectrophotometry and mass spectrum. The veterinary drug conjugates were immobilized on the polystyrene fluorescent microspheres/beads. There were competitive reactions between the veterinary drugs in the aqueous phase and that on the beads for combination with their specific biotinylated monoclonal antibodies. The optimum amount of the veterinary drug conjugates and the antibodies were optimized and selected. The detective standard curves were plotted. The specificity and the unknown samples were also determined by grouping according to different concentrations of the interferes and the samples. Meantime, the different microstructures of the surfaces of the beads were also observed by scanning electron microscope.
RESULTSCouplings were completed between small molecular veterinary drugs and BSA. The amounts of the three conjugates and the antibodies were optimized. The detective standard curves of the suspension array and their corresponding coefficients of determination (R2) were good (R2 > 0.99). The detection ranges of the three veterinary drugs were (40.00 - 6.25) x 10(5) ng/L, (50.00-7.81) x 10(5) ng/L and 1.00 x 10(3) - 7.29 x 10(5) ng/L respectively. Simultaneously, the specific detection of the suspension microarray was excellent and did not indicate significant cross-reactions. Errors between the found and the real are in the range of 8.09% - 17.03%. It can be considered that the relative standard deviations were relatively small. Successful couplings were also directly confirmed by the observation for microstructures of the surfaces of the beads by scanning of electron microscope and laid good foundation for the following responses.
CONCLUSIONThe high-throughput suspension microarray should provide a novel method for multi-analysis of the veterinary drugs and have a wide applicative prospects with simple operation, sensitive, rapid and low cost.
Chloramphenicol ; analysis ; Clenbuterol ; analysis ; Drug Residues ; analysis ; Estradiol ; analysis ; Microarray Analysis ; methods ; Veterinary Drugs ; analysis
3.Micro machining of micro-cantilever probes for efficient deposition for biochips.
Yong-Hai LI ; Gui-Fu DING ; Jun-Tao XU ; Yong-Hua ZHANG
Chinese Journal of Biotechnology 2005;21(2):227-232
Biochip technology will bring a tremendous revolution to life science and medical research in 21 century. Microarray assays represent an essential technical advance in biomedical research. Recently, the demand for microarray assay technology has spring up. Therefore, low cost and flexible techniques are needed to meet specific requirements for increasingly integrated biochips. Also performance must be improved in terms of speed and sensitivity. To this end, promising approaches, mainly based on micro and nanotechnologies, have been developed. In this paper, the design and microfabrication of a novel type of micro-cantilever probe are introduced. These probes were fabricated using silicon dioxide by Micro-electromechanical System (MEMS) techniques, and they featured one micron split gap, microchannels and self-replenishing reservoirs. All fabricated micro-cantilever probe were tested on Nanoarrayer instrumentation. Cy3-streptavidin was loaded as biological sample and patterned on DSU gold surface. Results showed these probes were capable of generating high quality biological arrays with routine spot sizes of 2 - 3 microns and could deposit at least three thousand spots without reloading. The spot size could potentially achieve sub-micron when probe size was further shrunk down by the high-resolution lithography technique or more precise microfabrication technologies, such as E-beam lithography. To further improve sample loading efficiency, it is needed to modify the cantilever surface in order to better confine sample inside the microchannel and reservoir, which will be researched in the future.
Microarray Analysis
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instrumentation
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methods
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Microelectrodes
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Molecular Probe Techniques
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instrumentation
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Nanotechnology
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instrumentation
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methods
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Silicon Dioxide
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chemistry
4.Application of chromosomal microarray analysis for a cohort of Chinese patients with spontaneous miscarriage.
Haiming YUAN ; Mengfan CHEN ; Xiaoyan DENG ; Fen LYU
Chinese Journal of Medical Genetics 2016;33(4):442-446
OBJECTIVETo explore the relationship between spontaneous miscarriage and chromosomal aberrations identifiable with chromosomal microarray analysis (CMA).
METHODSA total of 440 product-of-conceptions were collected for the CMA testing.
RESULTSFour hundred and seventeen of 440 specimens (94.7%) were successfully detected, among which 209 (50.1%) were chromosomal abnormalities. One hundred and twenty-nine (61.7%) of the 209 specimens were numerical chromosomal abnormalities, 40 specimens (19.1%) were structural anomalies, 38 specimens (18.1%) were mosaicisms, and 2 specimens (1.0%) showed regions of homozygosity.
CONCLUSIONCMA analysis of products of-conception specimens can yield a higher diagnostic rate than conventional karyotyping. The identification of the cause of spontaneous miscarriage can facilitate estimation of recurrence risks for future pregnancies.
Abortion, Spontaneous ; etiology ; genetics ; Chromosome Aberrations ; Cohort Studies ; Female ; Humans ; Karyotyping ; Microarray Analysis ; methods ; Pregnancy
5.Application of microfluidic-chip in biomedicine.
Chinese Journal of Biotechnology 2006;22(1):167-171
As a novel analytical technology, the research of Micro total analysis systems (micro-TAS) has been spreading rapidly. micro-TAS has been widely used to perform chemical and biochemical analysis. Microfluidic-based analytical system as micro-TAS's manily direction develops very fast in terms of it's reaction speed, reagent consumption, miniaturization, cost, and automation. After having proven the value of microfluidics for genetic, proteomic and cytomics analysis, this article also anticipates the development tendency of this technology in the biology medicine domain. It has demonstrated that a truly, easy-to-handle Microfluidic-based analytical device will be emerged in the future.
Humans
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Microarray Analysis
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instrumentation
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methods
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Microfluidic Analytical Techniques
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trends
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Microfluidics
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trends
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Technology Assessment, Biomedical
6.Chromosome microarray analysis of four fetuses with abnormal karyotypes.
Ruifang ZHU ; Xiangyu ZHU ; Ying YANG ; Xing WU ; Ying ZHANG ; Yiyan SHI ; Linlin HE ; Jie LI
Chinese Journal of Medical Genetics 2017;34(6):812-815
OBJECTIVETo carry out chromosomal microarray analysis (CMA) on four fetuses with abnormal karyotypes.
METHODSAmniotic fluid samples were obtained and subjected to routine G-banded karyotyping analysis. CMA was applied for cultured amniocytes to determine alterations of gene dosage and chromosomal breakpoints.
RESULTSAbnormal karyotypes were found in the parents of 3 fetuses. Parental karyotypes of the remaining fetus were normal. Imbalance chromosome rearrangements were revealed by CMA in all 4 cases.
CONCLUSIONCMA is an effective tool for the evaluation of clinical significance and delineation of the breakpoints involved in complex chromosomal rearrangements.
Abnormal Karyotype ; Adult ; Chromosome Banding ; Female ; Humans ; Karyotyping ; Microarray Analysis ; methods ; Pregnancy ; Prenatal Diagnosis
7.New strategies for the treatment of carcinoma of unknown primary.
Chinese Journal of Oncology 2023;45(1):44-49
Carcinoma of unknown primary (CUP) is a kind of metastatic tumor whose primary origin cannot be identified after adequate examination and evaluation. The main treatment modality of CUP is empiric chemotherapy, and the median overall survival time is less than 1 year. Compared with immunohistochemistry, novel method based on gene expression profiling have improved the sensitivity and specificity of CUP detection, but its guiding value for treatment is still controversial. The approval of immune checkpoint inhibitors and pan-cancer antitumor agents has improved the prognosis of patients with CUP, and targeted therapy and immunotherapy based on specific molecular characteristics are the main directions of future research. Given the high heterogeneity and unique clinicopathological characteristics of CUP, "basket trial" is more suitable for clinical trial design in CUP.
Humans
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Neoplasms, Unknown Primary/genetics*
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Carcinoma/drug therapy*
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Gene Expression Profiling/methods*
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Microarray Analysis
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Prognosis
8.Application of chromosome microarray analysis for fetuses with multicystic dysplastic kidney.
Feifei CHEN ; Tingying LEI ; Fang FU ; Ru LI ; Yongling ZHANG ; Xiangyi JING ; Xin YANG ; Jin HAN ; Li ZHEN ; Min PAN ; Can LIAO
Chinese Journal of Medical Genetics 2016;33(6):752-757
OBJECTIVETo explore the genetic etiology of fetuses with multicystic dysplastic kidney (MCDK) by chromosome microarray analysis (CMA).
METHODSSeventy-two fetuses with MCDK were analyzed with conventional cytogenetic technique, among which 30 fetuses with a normal karyotype were subjected to CMA analysis with Affymetrix CytoScan HD arrays by following the manufacturer's protocol. The data was analyzed with ChAS software.
RESULTSConventional cytogenetic technique has revealed three fetuses (4.2%) with identifiable chromosomal aberrations. CMA analysis has detected pathogenic CNVs in 5 fetuses (16.7%), which included two well-known microdeletion or microduplication syndromes, i.e., 17q12 microdeletion syndrome and Williams-Beuren syndrome (WBS) and three submicroscopic imbalances at 4q35.2, 22q13.33, and 1p33. PEX26, FKBP6, TUBGCP6, ALG12, and CYP4A11 are likely the causative genes.
CONCLUSIONCMA can identify the submicroscopic imbalances unidentifiable by conventional cytogenetic technique, and therefore has a significant role in prenatal diagnosis and genetic counseling. The detection rate of pathogenic CNVs in fetuses with MCDK was 16.7% by CMA. 17q12 microdeletion syndrome and WBS are associated with MCDK. Mutations of PEX26, FKBP6, TUBGCP6, ALG12, and CYP4A11 genes may be the causes for MCDK.
Adult ; Chromosomes ; genetics ; Female ; Fetus ; Humans ; Male ; Microarray Analysis ; methods ; Multicystic Dysplastic Kidney ; genetics ; Pregnancy ; Prenatal Diagnosis ; methods ; Young Adult
9.Application of chromosome microarray analysis for the delineation of pathogenesis for fetal ventriculomegaly.
Zhouzhou LI ; Fang FU ; Tingying LEI ; Ru LI ; Xiangyi JING ; Xin YANG ; Jin HAN ; Min PAN ; Li ZHEN ; Can LIAO
Chinese Journal of Medical Genetics 2017;34(4):576-582
OBJECTIVETo assess the value of genome-wide high-resolution chromosomal microarray analysis (CMA) for the delineation of pathogenesis for fetal ventriculomegaly diagnosed by ultrasound or magnetic resonance imaging (MRI).
METHODSThree hundred and forty-one cases of fetal ventriculomegaly were collected. The samples were grouped based on the extent of lateral ventricular dilatation, presence of additional features, site of occurrence, and the maternal age. All samples were subjected to karyotyping analysis. For those with a normal karyotype, DNA was extracted and hybridized with an Affymetrix CytoScan HD array. All cases were followed up.
RESULTSAmong the 341 fetuses, 21 (6.2%) were detected with an abnormal karyotype. For the 320 cases with a normal karyotype, 179 (55.9%) have accepted CMA analysis. Potentially pathogenic CNVs were identified in 12 (6.7%) of the 179 cases, whose sizes ranged from 198 kb to 8.71 Mb. These included a 1q21.3q23.1 deletion, a 2q37.3 deletion, a 3p14.1p13 deletion, a 6q25.3 deletion, a 8q11.23 duplication, a 10q21.1 deletion, a 15q11.2 deletion and a 16p13.11p12.3 duplication, a 22q13.33 duplication, a 22q11.21 duplication and a Xp21.1 duplication (Duchenne muscular dystrophy). Pathogenic CNVs were detected respectively in 7.5% and 3.1% of those with mild and severe ventriculomegaly (P=0.615), in 6.1% and 7.4% of those with isolated and non-isolated ventriculomegaly (P=0.732), in 5.6% and 7.9% of those with unilateral and bilateral ventriculomegaly (P=0.511), and in 6.7% of both elderly and non-elderly groups (P=1.000).
CONCLUSIONThe detection rate for abnormal karyotypes among fetuses with ventriculomegaly was 6.2%. CMA can increase the detection rate by approximately 6.7%. There was no significant correlation between ventriculomegaly and presence of pathogenic CNVs. In clinical practice, fetuses with ventriculomegaly and a normal karyotype should be considered for CMA analysis.
Adult ; Chromosome Aberrations ; Female ; Fetus ; abnormalities ; Humans ; Hydrocephalus ; genetics ; Karyotyping ; methods ; Microarray Analysis ; methods ; Pregnancy ; Prenatal Diagnosis ; Young Adult
10.Gene function and microbial community structure in sulfide minerals bioleaching system based on microarray analysis.
Li SHEN ; Xueduan LIU ; Guanzhou QIU
Chinese Journal of Biotechnology 2008;24(6):968-974
Biohydrometallergy technology received more and more attention because of its simple process, low cost and kind to environment, especially in dealing with low-grade and complex minerals. However, it is difficult to optimize microorganism species and process parameters in bioleaching procedure because of the lack of suitable bacteria and quantitative analysis methods at micro-level for bioleaching system. This has resulted in the low efficiency and poor yield of the target metal in bioleaching. With the development of microarray and bacteria conservation technology, solutions to the above problems were being found. This article summarizes the latest findings on genetic elucidation and the community structure of microorganisms in sulfide minerals bioleaching system, in the aim of providing a better understanding on the significance of cross-field technology of biohydrometallergy and genomics.
Acidithiobacillus
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genetics
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isolation & purification
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metabolism
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Industrial Microbiology
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methods
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trends
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Microarray Analysis
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Minerals
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metabolism
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Mining
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methods
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Sulfides
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chemistry
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metabolism