Frontiers in Microbiology
·
2021
·
IF 5.64
Wang Wenbin,Liu Jianxin,Guo Shanshan,Liu Lei,Yuan Qianyun,Guo Lei,Pan Saikun
The discovery of outer membrane proteins (OMPs) with desirable specificity and surface availability is a fundamental challenge to develop accurate immunodiagnostic assay and multivalent vaccine of pathogenic Vibrio species in food and aquaculture. Herein 101 OMPs were systemically screened from 4,831 non-redundant proteins of Vibrio parahaemolyticus by bioinformatical predication of signaling peptides, transmembrane (TM) α-helix, and subcellular location. The sequence homology analysis with 32 species of Vibrio spp. and all the non-Vibrio strains revealed that 15 OMPs were conserved in at least 23 Vibrio species, including BamA (VP2310), GspD (VP0133), Tolc (VP0425), OmpK (VP2362), OmpW (VPA0096), LptD (VP0339), Pal (VP1061), flagellar L-ring protein (VP0782), flagellar protein MotY (VP2111), hypothetical protein (VP1713), fimbrial assembly protein (VP2746), VacJ lipoprotein (VP2214), agglutination protein (VP1634), and lipoprotein (VP1267), Chitobiase (VP0755); high adhesion probability of flgH, LptD, OmpK, and OmpW indicated they were potential multivalent Vibrio vaccine candidates. V. parahaemolyticus OMPs were found to share high homology with at least one or two Vibrio species, 19 OMPs including OmpA like protein (VPA073), CsuD (VPA1504), and MtrC (VP1220) were found relatively specific to V. parahaemolyticus. The surface proteomic study by enzymatical shaving the cells showed the capsular polysaccharides most likely limited the protease action, while the glycosidases improved the availability of OMPs to trypsin. The OmpA (VPA1186, VPA0248, VP0764), Omp (VPA0166), OmpU (VP2467), BamA (VP2310), TolC (VP0425), GspD (VP0133), OmpK (VP2362), lpp (VPA1469), Pal (VP1061), agglutination protein (VP1634), and putative iron (III) compound receptor (VPA1435) have better availability on the cell surface.
研究领域:
微生物学
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分子生物学
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免疫学
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生物信息学
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食品科学
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水产养殖
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DOI
PubMed
高影响因子
JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY
·
2016
·Vol. 16
·期 3
·2291-2295
·
IF 1.338
Liu Daofeng,Yu Zhibiao,Huang Yanmei,Wang Shuying,Wang Jingyun,Guo Qi,Xu Chaolian,Xia Shiqi,Lai Weihua
A competitive lateral flow assay for the rapid detection of Salmonella choleraesuis was developed. Immuno-magnetic nanobeads were produced by covalently coupling anti-Salmonella choleraesuis antibody to magnetic nanobeads. These immuno-magnetic nanobeads were used as visually
detected probes in the subsequent assay. Compared with the traditional sandwich assay, which is used for detecting macro-molecules, this new method was developed based on the competitive relationship between S. choleraesuis in the inspected sample and the outer membrane protein immobilized
on the T line. Thus, only one antibody was necessary in the new assay, whereas a pair of rigorously selected antibodies were required in the sandwich assay. The sensitivity of the competitive assay for S. choleraesuis was 1.2×107 cfu/mL. In addition, no cross reactions were
found in the 17 common non-Salmonella bacteria strains and in the 4 Salmonella strains of other serotypes. Thus, with satisfactory sensitivity and specificity, the assay can be applied for the rapid detection of pre-enriched culture that may contain S. choleraesuis.
研究领域:
微生物学
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生物技术
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食品安全
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诊断检测
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Weixia Zhi,Xinjie Guo,Yichu Nie,Lu Nie,Tingting He,Li Dai,Jing Xu,Xiangxin Zong,Jun Xu,Huaihong Cai,Yong Tang,Yanguang Cong,Jiang Pi,Youfeng Zhu,Pinghua Sun,Jialiang Guo,Huoqiang Chen,Xueqin Huang,Haibo Zhou
The current lack of rapid screening methods for Klebsiella pneumonia ( K. pneumonia ) poses a considerable threat to human health. Herein, we fabricated a multilayer Au@Au@Ag/Pt nanoparticle (NP) that made of the peroxidase-mimicking external shell and tunable plasmonic intra-nanogap as a versatile probe, and conjugated them with lateral flow immunoassay (LFIA) for colorimetric-catalytic-SERS (CM/CL/SERS) multi-model sensing K. pneumonia in complex biological matrix. After the optimization of several parameters, the limit of detection (LOD) of the target K. pneumonia were 10 3 CFU/mL for CL-LFIA and 38 CFU/mL for SERS-LFIA, both of which were over 10-fold and 200-fold more sensitive than the visual CM-LFIA (10 4 CFU/mL), respectively. The clinical validation of this multi-model LFIA was performed with the infected patients ( N = 6) and healthy subjects ( N = 3), among which 4 positive samples missed by conventional CM-LFIA were successfully identified by SERS-LFIA. More importantly, this LFIA has great specificity for the target bacteria, while showed no signal response to samples infected by other bacteria. Different from previous studies focusing on detection of protein and nucleic acid of K. pneumonia , our study shortened the sample-to-answer time with direct detection of K. pneumonia by LFIA, filling the diagnostic gap for point-of-care testing (POCT) of K. pneumonia infection. Overall, the great performance of our established LFIA suggested its substantial potential for diagnosis of K. pneumonia -induced nosocomial infection.
研究领域:
纳米技术
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生物传感器
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分析化学
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生物医学工程
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微生物学
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诊断检测
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免疫学
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产品关联
DOI
高影响因子
Nature Protocols
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2025
·
IF 16
Lian Zewei,Chi Jimei,Yang Xu,Cheng Lijun,Xie Daixi,Tan Zhiyu,Chen Sisi,Yun Yang,Yibulayimu Yisilamu,Wu Wei,Song Yanlin,Su Meng
There is an urgent requirement to improve accessibility to diagnostic tools in remote areas. This requires assays that are easy to use, are cost-effective and produce rapid results. Important public health applications include early disease diagnosis, real-time monitoring, epidemic control and medical cost control. This protocol describes the fabrication of all-printed photonic crystal (PC) biochips for point-of-care testing of biomarkers. The photonic crystal material is prepared by the self-assembly of latex nanospheres that are printed onto a polyethylene terephthalate substrate. Photonic crystals composed of latex nanospheres of different sizes enhance the fluorescent signal emitted at different wavelengths, resulting in remarkably higher detection sensitivity. PC microarrays enable mass-printed preparation (up to 2,700 pieces can be printed per hour by one printer), and each microarray can be stored for a long time (>6 months) after heating. Biomarker specificity is achieved by the biofunctionalization of the nanospheres, for example, attaching capture antibodies. The detection involves the use of a fluorescently labeled detection antibody and a simple point-of-care detection device. This universal approach can be applied to the detection of many biomarkers, and the simultaneous detection of multiple biomarkers is also possible. Here we demonstrate describing how to prepare a chip that can be used to detect three inflammatory biomarkers in 10 min at low sample volumes at a cost of less than 3 CNY (~US$0.41) per PC codetection biochip. The biofunctionalization process including capture antibody coupling and blocking takes 3–4 h, and the detection process takes 20 min.
研究领域:
生物医学工程
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纳米技术
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诊断检测
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生物传感器
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公共卫生
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材料科学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Yuanzhe Li,Wenchen Gu,Yaling Li,Zheng Zhao,Ting Liu,Peifeng Ke,Haibiao Lin,Qiqin Wang,Xianzhang Huang,Qiaoxuan Zhang
Plasma brain-derived extracellular vesicles (BDEVs) possess excellent potential for the early diagnosis of Alzheimer's disease (AD). However, their accurate detection is often trapped by the low levels of specific BDEVs and the serious interference of high abundance surface proteins on EVs. In this study, an explosive signal amplification biosensing platform based on the novel “hedgehog-nanovesicles redox probe” was developed for the ultra-sensitive detection of plasma BDEVs. The novel probe was self-assembled by the target BDEVs, and it immobilized by methylene blue-conjugated aptamers, which enabled a dual-signal ratiometric sensor. Owing to the multi-amplification capability of novel probe and the built-in correction factor of dual-signal, the proposed platform exhibited higher sensitivity (with a limit of detection of 2.63 × 10 2 particles/mL), non-specific interferences, simpler operation, and more efficient preparation than previous methods. In the analysis of clinical samples, AD and mild cognitive impairment (MCI) patients can be differentiated from healthy individuals with AUC values of 0.823 and 0.833. The diagnostic accuracy of AD and MCI can be significantly improved with AUC values of 0.963 and 0.949 after combined with plasma Aβ42/Aβ40. It confirmed that BDEVs possess strong potential for AD early diagnosis, and particularly the sensor strategy provided a powerful tool for this purpose.
研究领域:
生物传感器
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纳米技术
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神经退行性疾病
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诊断检测
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生物分析化学
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分子诊断学
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DOI
高影响因子
ANALYTICAL LETTERS
·
2025
·
IF 1.8
Xiu Fan,Xiaoyan Li,Yihua Wang,Dawei Xu,Liping Zhao,Ruilan Yuan,Chonghua Hao,Chunyu Wang
Staphylococcus aureus (S. aureus) is globally recognized as a foodborne pathogen that causes food poisoning by secreting enterotoxins. In this study, a novel dual-recognition strategy for detecting S. aureus was developed, employing vancomycin-functionalized magnetic beads (Van-MBs) for target capture and canine immunoglobulin G (IgG) to selectively and sensitively bind to staphylococcal protein A (SPA) on the bacterial surface. Dual-recognition strategies based on vancomycin and porcine IgG have previously been developed for the detection of S. aureus. However, porcine IgG can also bind to protein G, which poses the risk of generating false-positive signals from protein G-producing Streptococci. In contrast, canine IgG demonstrates a higher affinity for protein A than for protein G. When S. aureus is present in the sample matrix, the Van-MBs and biotin-labeled canine IgG successfully form a sandwich complex. Then, streptavidin-conjugated horseradish peroxidase (HRP) is added, which binds to the sandwich structure. HRP then catalyzes the oxidation of 3,3’,5,5’-tetramethylbenzidine (TMB), resulting in the formation of a colorimetric signal that was proportional to the concentration of S. aureus within the concentration range of 6.3 × 103–4.0 × 105 CFU/mL with a detection limit of 1.9 × 103 CFU/mL. By substituting porcine IgG with canine IgG, this method eliminates cross-reactivity with protein G-expressing Streptococci, achieving specificity over non-target species. This approach highlights its potential for application in diverse sample matrices, including clinical and biological specimens.
研究领域:
微生物学
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免疫学
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食品卫生
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诊断检测
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纳米技术
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Xiaolong Guo,Yan Zhao,Xuemei Xu,Zhibin Wu,Li Xiao,Xiaoyang Ma,Ziqiu Lei,Yilin Zhao,Chengli Zhang,Lixing Song,Minggang Yin
Rapid and accurate detection of Porphyromonas gingivalis ( P. gingivalis ) is crucial for the timely diagnosis and management of periodontal diseases as well as for elucidating its role in systemic conditions. However, conventional diagnostic methods are complex, time-consuming, and unsuitable for point-of-care testing, necessitating the development of a more efficient and accessible detection system. This study presents a novel dual-probe fluorescence platform featuring cadmium telluride quantum dots (CdTe QDs) and a specially designed probe (PAN) as fluorescent molecules, which does not require a washing step. The PAN and CdTe QD probes can efficiently differentiate between Cu 2 + and Cu + ions through ionic hydrolysis and cation exchange reactions, respectively, resulting in the sensitive and rapid detection of arginine-specific gingipain, a key biomarker for P. gingivalis . Furthermore, the color change of CdTe QDs upon contact with saliva samples facilitates the visual identification of P. gingivalis . This dual-probe platform demonstrated a higher signal-to-noise ratio and greater reliability compared to the single-probe methods. Moreover, this platform features a low detection limit of 5 and 20 colony-forming units (CFUs)/mL for the PAN and CdTe QDs signals, respectively, with a visual detection threshold as low as 200 CFU/mL. Thus, this platform provides quantitative and qualitative results that are 100 % in concordance with quantitative PCR results from clinical saliva samples of patients with periodontitis and healthy controls. This simple, rapid, and cost-effective diagnostic tool can be potentially used for point-of-care applications, enabling early detection and improved monitoring of oral health and related diseases.
研究领域:
生物医学工程
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微生物学
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牙科医学
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生物传感器
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纳米技术
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诊断检测
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2025
·
IF 3.8
Wang Hongying,Li Zhiyan,Yang Wei,Chang Le,Yan Cunling
Conformational and colloidal stabilities are responsible for the stability of hemoglobin. However, most hemoglobin A1c (HbA 1c ) control materials (e.g., reference materials, calibrators, and quality control materials) are unstable and noncommutable. This may be closely related to the breakdown of the balance between the conformational and colloidal stabilities of hemoglobin. Therefore, we developed a new combined formulation buffer to address the causes of conformational and colloidal instabilities and to maintain hemoglobin or HbA 1c stability. For the commutability of HbA 1c control materials, we purified hemoglobin to remove plasma proteins, phospholipids, and fats. On the basis of the above design, we prepared two medical concentrations of HbA 1c control materials for the external quality assessment (EQA) program and then determined the certified values via the reference measurement method. Stability, homogeneity, value transfer, and commutability were evaluated. Moreover, the control material was distributed to 56 laboratories to assess proficiency testing. The HbA 1c control materials showed good homogeneity and stability and reliable value transfer, and were commutable for routine clinical analyzers. In the EQA program, only one analyzer exceeded the acceptable bias range (± 6.0%), and the mean interlaboratory CV s of the three analyzers exceeded the acceptable range (3.0%), which can be used to assess true values to improve proficiency testing in the EQA program. Therefore, newly developed control materials can be used to standardize the determination of HbA 1c . Graphical
研究领域:
临床化学
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诊断检测
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实验室医学
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生物化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2025
·
IF 3.8
Wang Chengxue,Wang Qian,Yuan Wenting,Jia Zichao,Dong Lizhen,Chai Yamin,Cai Qinzhen,Yuan Chunhui,Cao MingFeng,Luo Wei
Dichloromethane (DCM), a traditional extractant for 24-h urine free cortisol (UFC) testing, poses significant occupational hazards. This study aimed to explore a novel extractant, evaluate its performance in replacing DCM, and establish the biological reference range for UFC detection using the new extractant. Comparative analyses of extraction efficiency were conducted between ethyl acetate (EA) and DCM. A stabilized EA-based formulation (named EEG) was developed by incorporating anhydrous ethanol and gallic acid as anti-hydrolysis and volatility-reducing agents. Recovery rates were assessed using a control product. Stability was tested through delayed detection. Biological reference intervals were determined via chemiluminescence immunoassay (CLIA) and liquid chromatography-tandem mass spectrometry (LC–MS/MS) following EEG extraction, and determined the reference interval using mean ± 1.96 standard deviation. EA demonstrated superior extraction efficiency, yielding significantly higher median cortisol levels (6.27 (IQR 4.68–8.68) µg/dL) versus DCM (3.96 (IQR 2.41–5.07) µg/dL, p = 0.002). Recovery tests confirmed EA’s enhanced performance (mean recovery 95.3% vs. DCM’s 65.4%, p = 0.011). The EEG formulation exhibited improved stability with unchanged extraction efficacy. Established reference intervals (95% range) were 14.58~136.62 µg/24 h (CLIA) and 0.81~60.55 µg/24 h (LC–MS/MS). EA extraction surpasses DCM in recovery and safety. The optimized EEG formulation demonstrates enhanced stability while maintaining analytical reliability. The newly established reference intervals provide method-specific benchmarks for clinical UFC assessment. This innovation offers a safer, more efficient alternative for routine laboratory practice. The findings underscore the feasibility of replacing hazardous solvents in endocrine diagnostics without compromising analytical performance.
研究领域:
临床化学
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内分泌学
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分析化学
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诊断检测
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职业健康
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 8
·1-12
·
IF 5.3
Ji Jingjing,Li Jiahao,Wu Minghui,Wang Bin,Lu Yu-nan,Zhang Chuhan,Shan Yanke,Liu Fei
A multiplex colloidal gold immunochromatographic assay (mCGIA) was developed for the simultaneous detection of feline calicivirus (FCV), feline panleukopenia virus (FPV), and feline herpesvirus (FHV) antibodies in feline serum. The expressed and purified recombinant FCV-VP1, FPV-VP2, and FHV-gD proteins exhibited strong antigenic reactivity. The mCGIA test strip achieved rapid (< 15 min) and specific detection, showing no cross-reactivity with other feline viral antibodies. The assay achieved a low visual limit of detection (vLOD) of 1:1 × 10 5 for FCV and 1:1 × 10 4 for FPV and FHV antibodies. Stability testing revealed 90% retention of detection capability after 4 weeks of storage at 37 °C. Reproducibility was robust, with intra- and inter-assay coefficients of variation (CVs) of less than 7% and 13%, respectively. The smartphone-based portable reader enabled quantitative detection with wide linear ranges (from 1:1000 to 1:100000 for FCV, 1:250 to 1:10000 for FPV and FHV) and excellent linearity (FCV: R 2 = 0.992, FPV and FHV: R 2 = 0.991). Compared with indirect ELISA using 164 clinical serum samples, the assay showed 98.17% (FCV), 98.78% (FPV), and 97.56% (FHV) concordance rates, respectively. This novel mCGIA platform provides a rapid, cost-effective tool for sero-surveillance, vaccine evaluation, and assisting clinical diagnosis, enhancing feline health management. Graphical
研究领域:
兽医学
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免疫学
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诊断检测
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病毒学
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即时检测
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生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）