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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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Endocrine-disrupting chemicals in aquatic ecosystems and their impacts on aquatic organisms and humans with advances in detection and remediation 认领 引用
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作者 Tariq DILDAR Yongcheng XUE +3 位作者 Wenxiao CUI Mhd IKHWANUDDIN Muhammad SHAFIQ Hongyu MA 《Journal of Zhejiang University-SCIENCE B》 SCIE CAS CSCD 2026年第6期561-589,I0002-I0012,共29页
The increasing anthropogenic burden,driven by population growth and intensified industrial and agricultural practices,has led to the widespread release of endocrine-disrupting chemicals(EDCs)into aquatic ecosystems,wi... The increasing anthropogenic burden,driven by population growth and intensified industrial and agricultural practices,has led to the widespread release of endocrine-disrupting chemicals(EDCs)into aquatic ecosystems,with significant implications for both environmental and human health.Many studies have reported the concentrations and toxicological effects of EDCs in aquatic environments,but few have addressed detection methods and remediation techniques.This review aims to highlight the sources,dynamics,and bioaccumulation of EDCs in aquatic ecosystems,along with their toxic effects on aquatic species and associated health risks in humans.Additionally,we provide an overview of advanced detection and remediation techniques.Our review found that EDCs,particularly phthalates and bisphenols,included in industrial effluents,domestic waste,and agricultural runoff,are frequently discharged into aquatic bodies through human activities.EDCs are associated with various toxic effects in aquatic organisms,such as bioaccumulation,transgenerational effects,reduced growth,immunotoxicity,DNA damage,and abnormal hormonal release,which impair reproductive development.Among the detection methods,biosensors,surface-enhanced Raman spectroscopy(SERS),and nuclear magnetic resonance(NMR)spectroscopy are promising tools for EDC detection relative to conventional analytical methods in aquatic systems.Emerging remediation techniques,such as hybrid activated carbon systems and N-doped carbon-based adsorbents,are recommended for their high removal efficiency.This review serves as a valuable resource for advancing research on EDC toxicity,detection,and remediation technologies. 展开更多
关键词 Endocrine-disrupting chemical Bioaccumulation Health risk Biosensor-based detection Hybrid activated carbon N-doped carbon adsorbent
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