ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Ling Deng,Xiaoyi He,Ke Liu,Yuanli Li,Han Xia,Hang Qian,Xiaoxue Lu,Xuhu Mao,Yang Xiang
Burkholderia pseudomallei is the causative agent of melioidosis, a potentially life-threatening infectious disease, and poses public health risks in endemic areas. Due to the high mortality, intrinsic antibiotic resistance , and atypical manifestations, establishing a rapid, accurate, and sensitive identification of B. pseudomallei enables earlier diagnosis, proper treatments, and better outcomes of melioidosis. Herein, we present a O ne- P ot C RISPR-integrated assay for I nstant and V isual D etection (termed OPC-IVD) of B. pseudomallei . The integration of recombinase polymerase amplification and CRISPR-Cas12a recognition-activated trans -cleavage, achieved a true all-in-one single-tube reaction system, initiating the amplification and cleavage simultaneously, which realized a facile sample-to-answer assay. This approach could be performed with simplified DNA extraction and completed around 30 min by holding the reaction tube in the hand. The detection limit of our OPC-IVD was determined to be 2.19 copy/uL of plasmid DNA, 12.5 CFU/mL of B. pseudomallei , and 61.5 CFU/mL of bacteria in spiked blood samples, respectively. Furthermore, the introduction of internal amplification control effectively reduced the occurrence of false negatives, which was incorporated in the reaction system, and amplified simultaneously with the target and read by naked eyes. The assay exhibited 100% accuracy when evaluated in clinical isolates and samples. The streamlined workflow of our OPC-IVD of B. pseudomallei enables a field-deployable, instrument-free, and ultra-fast approach that can be utilized by non-expert personnel in the field of molecular diagnosis of melioidosis especially in under-resourced setting.
研究领域:
医学诊断
·
分子生物学
·
传染病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2022
·Vol. 189
·期 9
·1-10
·
IF 6.408
Wang Yuexin,Chen Rui,Shen Bo,Li Cai,Chen Junman,Wang Yanshuang,Tian Shen,Li Xuemiao,Luo Nini,Liu Rui,Ding Shijia,Zhu Chuanlong,Xia Qianfeng
A multifunctional catalytic nanomaterial (Co-MOF@AuNP@ABEI) composed of cobalt-doped metal–organic frameworks (Co-MOF), gold nanoparticles (AuNP), and N -(4-aminobutyl)- N -(ethylisoluminol) (ABEI) is reported. Co-MOF@AuNP@ABEI exhibits high synergistic and zero-distance catalytic properties, which are beneficial to the improvement of the detection sensitivity of an electrochemiluminescent (ECL) biosensor. After coupling with the ECL system and 3D magnetic walking nanomachine amplification strategy, the Co-MOF@AuNP@ABEI can achieve an ultrasensitive ECL assay of Burkholderia pseudomallei with the limit of detection (LOD) of 60.3 aM, which is 2 and 4 orders of magnitude lower than individual ECL system without the nanomachine (4.97 fM) and individual walking nanomachine (340 fM), and superior to the pathogenic bacteria analyses in the previous report. Moreover, the LOD of the proposed ECL detection system for the determination of B. pseudomallei in serum sample was as low as 9.0 CFU mL −1 . The relative standard deviations (RSD) of ECL intensity for the detection of five B. pseudomallei -spiked serum samples were 4.02%, 0.84%, 0.84%, 1.55%, and 0.21%, respectively. The recoveries of the ECL biosensor for the detection of B. pseudomallei DNA-spiked serum samples were 93.63 ~ 107.83%. Therefore, this work demonstrated that the developed multifunctional catalytic nanomaterial with synergistic and zero-distance catalytic properties can be used as excellent ECL signal reporter to improve the detection sensitivity of ECL biosensor. Graphical abstract
研究领域:
生物传感器
·
分析化学
·
纳米技术
·
微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
iScience
·
2026
·Vol. 29
·期 4
·
IF 4.5
Chuizhe Chen,Xuemiao Li,Xin Li,Kangji Yang,Bo Wang,Lianpan Dai,Qianfeng Xia
Burkholderia pseudomallei ( B. pseudomallei ) exhibits inherent resistance to multiple antibiotics and Strong pathogenicity, highlighting the urgent need for effective antibiotics. The type VI secretion system (T6SS) is a key virulence factor in B. pseudomallei , and its core structural protein hemolysin coregulated protein (Hcp) represents a conserved, essential target for potential intervention. In this study, Hcp was utilized as a molecular target to screen antibacterial agents from the FDA Orange Book database. Subsequent functional assays and mechanistic analyses identified netilmicin sulfate, which specifically binds to Hcp and exhibits a potent bacteriostatic effect. In vitro and in vivo studies demonstrated that it effectively eliminates B. pseudomallei and enhances host survival post-infection. Combination with ceftazidime exhibits synergism, strengthening host cell protection and reducing the required dosage. Mechanistically, it disrupts Hcp’s tubular structure, compromising T6SS functional architecture and attenuating B. pseudomallei viability. Beyond identifying a promising antibiotic candidate, this approach underscores a potential strategy for future antimicrobial drug discovery targeting T6SS components.
✓ 已验证
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Yuexin Wang,Bo Shen,Nini Luo,Cai Li,Haiping Wu,Yanshuang Wang,Shen Tian,Xuemiao Li,Rui Liu,Xinmin Li,Junman Chen,Wei Cheng,Shijia Ding,Rui Chen,Meifang Xiao,Qianfeng Xia
The simple, rapid, and accurate detection of highly lethal melioidosis is crucial for early clinical diagnosis and improving cure rates. Currently, due to time consumption, low sensitivity and detection rate the existing clinical detection methods cannot satisfy the needs of clinical diagnosis. Herein, a novel self-enhanced porous hydrogel material (Au@PEI-ABEI@Pt) for ultrasensitive ECL strategy of detection B. pseudomallei was report. The novel porous hydrogel material composed of PEI-ABEI porous hydrogel, gold nanoparticles (AuNP) and platinum nanoparticles (PtNP) have large specific surface area and porous structure, which not only fix more ABEI to realize self-enhanced ECL signal amplification, but also facilitate ion diffusion and efficient utilization of catalytic materials, realizing rapid electron transfer and zero-distance catalysis, significantly improving the initial signal of ECL sensor. Moreover, the most attractive aspect is that Au@PEI-ABEI@Pt hydrogels with good biocompatibility can achieve widespread application of CRISPR/Cas12a in solid-phase carriers without affecting the sensitivity, specificity and shearing activity of CRISPR/Cas12a. After coupling with the ECL system and CRISPR/Cas12a signal amplification strategy, the Au@PEI-ABEI@Pt can achieve an ultrasensitive ECL assay of B. pseudomallei with the LOD of 5 CFU mL −1 in complex samples, with high specificity and stability to effectively classify B. pseudomallei and other Gram-negative bacteria. This study shows that the developed porous hydrogel materials not only serve as an excellent ECL signal reporter to significantly improve the detection sensitivity of ECL biosensors, but also provide a new approach for the wide application of CRISPR/Cas systems in solid-phase carriers.
研究领域:
生物技术
·
纳米技术
·
分析化学
·
生物传感器
·
分子生物学
·
医学诊断
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）