Animals
·
2023
·Vol. 13
·期 20
·3177
·
IF 3
Hongbo Zhang,Nan Zhang,Jianhua Li,Panpan Zhao,Xin Li,Xiaocen Wang,Xu Zhang,Bao Yuan,Fei Gao,Pengtao Gong,Xichen Zhang
Simple SummaryTritrichomonas muris(T. muris) mainly parasitizes the ceca of different rodent species worldwide. To date, there are no ideal methods for the detection ofT. murisinfections in laboratory mice. This study developed a nested PCR reaction using a novel specific primer based on the conserved regions of the SSU rRNA gene ofT. murisfor a molecular epidemiological survey (investigation) ofT. murisinfections in laboratory mice. The results showed that the nested PCR system has higher sensitivity, reliability, and specificity. The nested PCR system showed an infection rate ofT. murisof 18.96% (58/306), which was higher than the infection rate of 14.05% (43/306) that was detected via smear microscopy in fecal samples from five mouse strains. The sensitivity and specificity of nested PCR in detectingT. muriswas found to be 100%, and it demonstrated a 26% increase in diagnostic sensitivity compared to the smear microscopy method. The present study provides a new method for the molecular epidemiological investigation ofT. murisinfections in laboratory mice.AbstractA variety of rodent ceca are parasitized byTritrichomonas muris(T. muris), a flagellated protozoan. To date, there are no ideal methods for the detection ofT. murisinfections in laboratory mice; thus, new molecular methodologies for its specific detection need to be developed. In this study, using staining and SEM, it was observed thatT. murishas a pear-shaped body and contains three anterior flagella. A nested PCR system with novel specific primers was designed based on the conserved regions of the SSU rRNA gene ofT. muris. The nested PCR system forT. murisshowed good specificity and high sensitivity for at least 100T. muristrophozoites/mL and 0.1 ng/μL of fecal genomic DNA, which means that 176 trophozoites per gram of mouse feces could be detected. When using this nested PCR system, the detection rate was 18.96% (58/306), which was higher than the detection rate of 14.05% (43/306) detected via smear microscopy in fecal samples from five mouse strains. The sensitivity and specificity of nested PCR in detectingT. muriswas found to be 100%, and it demonstrated a 26% increase in diagnostic sensitivity compared to the smear microscopy method in the present study. In conclusion, the nested PCR developed with novel primers based on the SSU rRNA gene ofT. murishas good accuracy, specificity, and sensitivity for the detection ofT. murisinfections in laboratory mice.Keywords:Tritrichomonas muris;SSU rRNA;primer;nested PCR;laboratory mice
研究领域:
分子生物学
·
寄生虫学
·
兽医学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Chenguang Wang,Nan Cheng,Longjiao Zhu,Yuancong Xu,Kunlun Huang,Pengyu Zhu,Shuifang Zhu,Wei Fu,Wentao Xu
A colorimetric biosensor for DNA screening was designed based on the conformational changes of the horseradish peroxidase (HRP)-mimicking DNAzyme. The scheme of DNA biosensing was designed based on the base pairing of DNAzyme sequence to inhibit the formation of HRP-mimicking hemin/G-quadruplex structures in the process of amplification. DNA could be amplified via the universal primer multiplex polymerase chain reaction (UP- M -PCR) and innovatively detected as color disappear in the reaction visible to the naked eye. The input of key factors and the output of optical characteristics in the reaction inspired the development of an OR logic gate operation for DNA detection. This biosensor overcomes self-inhibition and amplification disparity with the help of UP- M -PCR, thereby exhibiting high specificity and high-throughput without the requirement of gel analysis work. This biosensing system also presented 1% sensitivity and approximately 180 copy numbers in triplicate. The biosensor was used to screen elements from genetically modified organisms (GMOs) and covered more than 90% of all globally authorized events in the world. The designed colorimetric biosensor is a rapid, portable and versatile tool for nucleic acids detection and diagnosis in the field.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
Nature Protocols
·
2025
·
IF 16
Xu Ziye,Lyu Yuexiao,Chen Haide,Chen Yu,Wang Yongcheng
Formalin-fixed paraffin-embedded (FFPE) samples represent a vast and valuable resource of patient material, often linked to extensive clinical history and follow-up data. However, achieving single-cell or single-nucleus RNA (sc/snRNA) profiling in these archived tissues remains challenging. To address this, we have developed snRandom-seq, a droplet- and random primer–based single-nucleus total RNA sequencing technology specifically designed for FFPE tissues. This method captures total RNAs by using random primers and demonstrates a low doublet rate (0.3%), increased RNA coverage and enhanced detection of non-coding and nascent RNAs compared to state-of-the-art high-throughput sc/snRNA-seq technologies. This protocol provides a comprehensive guide to isolating single nuclei from FFPE samples; performing in situ DNA blocking, reverse transcription and dA tailing reactions; barcoding single-nucleus droplets; and preparing sequencing libraries. The entire snRandom-seq process can be completed in 4 d. This platform serves as a powerful tool for snRNA-seq of clinical specimens, with broad applications in studying complex biological systems.
研究领域:
分子生物学
·
医学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Yue Guo,Shenglin Sun,Mingfei Gu,Liyue Zhang,Li Cheng,Zhaofeng Li,Caiming Li,Xiaofeng Ban,Yan Hong,Zhengbiao Gu
Linear α-glucan (LG), a linear polymer linked by α-1,4 bonds, has received increasing attention for its potential applications in synthetic polymer production. Notably, the functionality of LG is strongly influenced by its degree of polymerization (DP). In this study, SP and GP were successfully constructed and expressed. The reaction of enzymatic co -polymerization into LG was investigated. The preferred reaction was carried out at 37 °C and pH 7.4 for 72 h, with a maximum conversion rate of 25 %. Afterwards, two approaches were used to modulate the molecular structures of LGs. Firstly, LGs with distinct molecular weights ranging from 1062.33 ± 16.04 g/mol to 5679 ± 80.29 g/mol were obtained by adjusting the substrate/primer ratio during the LG synthesis process. Secondly, two distinct products could be produced by altering the enzyme addition method: short-chain LG with a DP < 10 (64.34 ± 0.54 %) or long-chain LG with a DP > 45 (45.57 ± 2.18 %). Additionally, theoretical synthesis model was constructed which subdivided the reaction into three stages to evaluate this dual-enzyme cooperative system. These findings have significant implications in promoting the application of LG in the fields of biomedicine and material science.
研究领域:
生物化学
·
材料科学
·
生物医药
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF DENTISTRY
·
2025
·
IF 5.5
Huimin Wang,Wei Wei,Qi Lin,Zilu Tian,Shuwei Qiao,Tingting Bai,Xi He,Song Zhu
Objectives The effects of different interface treatments on the interfacial bonding of inorganic fillers in composite resins and the overall performance of the composite resins were investigated. Methods Synthetic mussel derivatives (N-3,4-dihydroxyphenethyl methacrylamide, DMA) were used as primers to treat the inorganic filler bonding interfaces, and the effects of different concentrations of DMA on the shear strength of the bonding interface were tested. Silica nanoparticles were coated on the surface of the filler via the sol-gel growing, and the mechanical properties and aging resistance of the composite resin were tested. Results The DMA primer concentration of 0.1–1mg/mL substantially improved the shear strength of the bonding interface, and combined with the coating of silica nanoparticles, it improved the compressive strength and toughness of the aged composite resin. Conclusions This study indicated that the use of the DMA primer and coated silica nanoparticles increased the bonding strengths and stabilities at the organic-inorganic interfaces, and also improved the mechanical strengths and durability of the composite resins. Clinical significance Fortified with DMA and silica nanoparticles, the composite resin exhibits great bonding strength, hydrolysis resistance, and toughness. This innovation could stabilize the resin for long-term and lower the risk of secondary caries. This study paves the way for the clinical application of novel resins to prevent secondary caries in the future.
研究领域:
材料科学
·
牙科医学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2022
·
IF 5.895
Jialin Liu,Yanli Wang,Xiaoxiao Li,Zhengyu Jin,Birte Svensson,Yuxiang Bai
Glycogen-like glucan (GnG) is a unique hyperbranched polysaccharide nanoparticle which is drawing increasing attention due to its biodegradability and abundant short branches that can be functionalized. Because starch and GnG are both composed of glucose residues and have similar glucosidic bonds, GnG could be fabricated by sucrose phosphorylase, α-glucan phosphorylase, and branching enzymes from starch primers and sucrose. In this study, high-amylose starch, normal starch, and waxy corn starch were used as primers to synthesize GnG, and their impact on the fine structure of GnG was investigated. Structural analysis indicated that with increasing content of amylopectin in the starch primer, the proportion of short chains in GnG decreased, and the degree of β-amylolysis and α-amylolysis was enhanced. Amylose in the primer contributed to a compact and homogeneous structure of GnG, while amylopectin triggered the formation of branch points with a more open distribution. These findings provide a new strategy for regulating the fine structure of GnG.
研究领域:
生物化学
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 8
·1-9
·
IF 5.3
Zhou Yanmei,Che Shengjun,Wang Zhili,Zhang Xiaoru,Yuan Xunyi
An electrochemical sensor assisted by primer exchange reaction (PER) and CRISPR/Cas9 system (PER-CRISPR/Cas9-E) was established for the sensitive detection of dual microRNAs (miRNAs). Two PER hairpin (HP) were designed to produce a lot of extended PER products, which could hybridize with two kinds of hairpin probes modified on the electrode and initiate the cleavage of two CRISPR/Cas9 systems guided by single guide RNAs (sgRNAs) with different recognition sequences. The decrease of the two electrochemical redox signals indicated the presence of dual-target miRNAs. With the robustness and high specificity of PER amplification and CRISPR/Cas9 cleavage system, simultaneous detection of two targets was achieved and the detection limits for miRNA-21 and miRNA-155 were 0.43 fM and 0.12 fM, respectively. The developed biosensor has the advantages of low cost, easy operation, and in-situ detection, providing a promising platform for point-of-care detection of multiple miRNAs. Graphical abstract
研究领域:
生物化学
·
分子生物学
·
分析化学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Aggregate
·
2024
·
IF 13.9
Zhenxin Yang,Jiale Su,Junzhan Wang,Xuanhe Li,Fushun Li,Juntao Hu,Nan Chen,Zhang Tao,Delong Yang,Deng-Ke Wang,Qiang Zhu,Yuhui Liao,Zheng-Hong Lu
Solution coating of organic semiconductors offers great potential for achieving low-cost and high-throughput manufacturing of large-area and flexible electronics. However, the solution processability of semiconducting small molecules for fabricating uniform and reliable thin-film devices poses challenges due to the low viscosities of small-molecule solutions. Here, we report a universal approach employing a primer template (PT) to enhance the spreadability of small-molecule solutions on silicon wafers, enabling the spin-coating fabrication of uniform thin films composed of millimeter-scale grains with complete large-area coverage and well-ordered molecular packing. Using PT, we fabricated organic thin-film transistors (OTFTs) using solutions containing various small molecules such as rubrene and 2-decyl-7-phenyl-[1]benzothieno[3,2-b][1]benzothiophene. The device yield of all fabricated OTFTs is consistently 100% while achieving a high average mobility of 1.62 cm 2 V −1 s −1 with a device-to-device variation of 7.7% measured in ambient air condition. In addition, the utilization of PT resulted in a batch-to-batch variation of 12.5% in device performance over dozens of OTFT devices. The key industrial manufacturing metrics, such as device yield, reproducibility, and performance uniformity of the PT OTFTs, are among the best for devices fabricated using solution spin-coating techniques.
研究领域:
材料科学
·
电子工程
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2024
·
IF 4.1
Yian Hu,Chengtao Xu,Chao Zhao,Hong Liu
We report an electrochemical aptamer-based (E-AB) sensor for label-free quantitative detection of thrombin. In-situ synthesis of high-density aptamer replicas on the electrode is accomplished based on primer exchange reaction (PER), by which a DNA primer is continuously extended with the presence of a catalytic hairpin template and DNA polymerase. The PER enables the synthesis of long single-stranded DNA with replicated aptamer sequence for thrombin binding. The PER products on the surface of a gold electrode increase the density of aptamers on a limited electrode area, which can provide more effective binding sites for thrombin and therefore improve the performance of the sensor based on electrochemical impedance spectroscopy. Using our method, thrombin is quantitatively detected from 1.0 pM − 50 nM with a detection limit of 0.91 pM. Compared with the conventional E-AB sensor based on a 15-mer aptamer immobilized on the electrode, the detection limit and linear range of our method is improved. Therefore, we believe our method is promising for label-free electrochemical detection of biomarkers.
研究领域:
生物化学
·
分析化学
·
生物传感技术
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2020
·
IF 2.877
Fangyuan Zheng,Sufang Li,Sunan Wang,Tao Feng,Zejun Jiang,Jiarong Pan
Adulteration of high-quality meat with their cheaper counterparts can be minimized by rapid and reliable methods for detecting meat species. Here an isothermal cross-primer amplification (CPA) technique combined with colloidal gold nucleic acid test strips (CPA strips) was developed to differentiate cow, sheep, arctic fox , and pig meat. A simple primer design for multiplex differentiation using a universal single-labeled CPA primer system and four detection-level species-specific labeling primers were analyzed by colloidal gold-based test strip assay. Moreover, simultaneous detection of fox and pig meat on a double-test line strip was feasible. The CPA strip assay indicated a lower amounts sensitivity of 0.3 ng DNA when one targeted species was tested and a detection limit of 1% when arctic fox meat was detected in the meat mixtures. Using a minimal set of primers, this study provides a promising tool for detecting the species of different types of meat using a constant temperature amplification technology.
研究领域:
食品科学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ACS Nano
·
2024
·
IF 15.8
Zhipan Wang,Aimin Ma,Yiping Chen
Multiple and ultrasensitive detection of pathogenic bacteria is critical but remains a challenge. Here, we introduce a digital assay for multiplexed and target DNA amplification-free detection of pathogenic bacteria using botryoidal-like fluorescent polystyrene dots (PS-dots), which were first prepared through the hybridization reaction between primer exchange reaction chains and polystyrene nanospheres that encapsulated polymer dots for signal preamplification. The pathogenic bacteria’s DNA was cleavaged by the argonaute (Ago) protein-mediated multiple and precise cleavage reactions, where the obtained target sequences bridged the magnetic beads (MBs) and botryoidal-like PS-dots via a hybridization reaction, and the fluorescent MB-botryoidal PS-dot complexes were utilized as digital probes based on colors and sizes for digital encoding. An artificial-intelligence-fluorescent microsphere counting algorithm was applied to identify and count the fluorescent MBs for digital readout. This digital assay combined the ultrabright botryoidal-like PS-dots with Clostridium butyricum Ago’s precise enzyme cleavage properties, achieving simultaneous detection of three pathogenic bacteria with a linearity range from 102 to 106 CFU/mL without target DNA amplification within 1.5 h. This digital assay has also been applied to detect aquatic and clinical samples with accepted accuracy (98%), which offers an avenue for a next-generation multiplexed digital platform for pathogenic bacteria analysis.
研究领域:
生物技术
·
医学检验
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Weipeng Liu,Minjun Zhu,Hongxing Liu,Jitao Wei,Xiaoming Zhou,Da Xing
Searching for a strategy to enhance the efficiency of nucleic acid amplification and achieve exquisite discrimination of nucleic acids at the single-base level for biological detection has become an exciting research direction in recent years. Here, we have developed a simple and universal primer design strategy which produces a fascinating effect on isothermal strand displacement amplification (iSDA). We refer to the resultant primer as “invading stacking primer (IS-Primer)” which is based on contiguous stacking hybridization and toehold-mediated exchange reaction and function by merely changing the hybridization location of the primer. Using the IS-Primer, the sensitivity in detecting the target miR-21 is improved approximately five fold compared with the traditional iSDA reaction. It was further demonstrated that the IS-Primer acts as an invading strand to initiate branch migration which can increase the efficiency of the untwisting of the hairpin probe. This effect is equivalent to reducing the free energy of the stem, and the technique shows superior selectivity for single-base mismatches. By demonstrating the enhanced effect of the IS-Primer in the iSDA reaction, this work may provide a potentially new avenue for developing more sensitive and selective nucleic acids assays.
研究领域:
分子生物学
·
核酸化学
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）