MICROBIAL PATHOGENESIS
·
2024
·
IF 3.3
Zhikun Zhang,Dawei Shi,Haiwei Dou,Ruijie Wan,Qing Yuan,Peng Tu,Deli Xin
In previous study, lower levels of serum GP130 were reported in children with MPP. GP130 is an important signal transducer, the down regulation of which may influence host immune responses. In this study, we aimed to analyze the regulatory mechanism of GP130 during MP infection. Firstly, the mRNA and protein levels of GP130 both decrease and then increase with increasing multiplicity of infection (MOI: 1 to 40) of MP. The lowest levels of GP130 were detected at MOI of 5. Then, heat treated MP but not trypsin treated MP or MP extracted proteins show regulatory effect to the expression of GP130. These indicate that the down regulation of GP130 is related to protein mediate adhesion process of MP. Gene expression analysis revealed that MP affected apoptosis and the TLR4 pathway in infected cells, and the mRNA level of IL-6 was correlated with that of GP130. Further, Z-VAD-FMK (pan-caspase inhibitor) can suppress the apoptosis induced by MP infection and restore GP130 at protein level. Further studies revealed that MP infection promoted TLR4 internalization but did not activate the NF-κB pathway. The levels of surface TLR4 showed correlation with the transcription of IL-6 and GP130. TAK242 (TLR4 inhibitor) and PS341 (proteasome inhibitor) can restore the decreased transcription of GP130, both of which were able to promote NF-κB pathway activation in MP-infected cells. These suggested that the regulation of TLR4/NF-κB pathway and induced apoptosis post MP infection are involved in the down-regulation of GP130 at transcription and protein levels, respectively.
研究领域:
微生物学
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免疫学
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细胞生物学
·
分子生物学
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DOI
PubMed
高影响因子
Sensors and Actuators Reports
·
2025
·
IF 7.6
Xiaopeng Chen,Wentao Wang,Pin Zhang,Qionglin Wang,Kangbo Liu,Fengfang Ding,Xianwei Zhang,Ligong Hou,Yingyu Zhang,Wancun Zhang
Simple, cost-effective, sensitive, and specific approaches that are capable of enabling point-of-care testing (POCT) are crucial for clinical diagnosis, especially in settings with extremely limited resources. Therefore, a universal, simple, low cost, sensitive, and specific POCT approach called RCA-F-LFS was developed by combining molecular beacon (MB)-assisted rolling circle amplification (RCA) for target amplification with fluorescent lateral flow strips (F-LFS) to enable highly sensitive and visual detection of RCA products. Using Mycoplasma pneumoniae as the detection target, RCA-F-LFS demonstrated high sensitivity (0.1 pM) and specificity, accurately distinguishing it from other pathogenic microorganisms. In addition, RCA-F-LFS enables visual detection of both Mycoplasma pneumoniae DNA and RNA. The detection results of clinical throat swab samples showed that RCA-F-LFS could accurately detect Mycoplasma pneumoniae , with 100 % agreement with current clinical approaches. As a proof of concept, RCA-F-LFS can detect other nucleic acid targets by simply changing the target sequence, offering a robust and versatile strategy for the development of novel clinical POCT detection methods.
研究领域:
分子诊断
·
即时检测
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核酸扩增
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生物传感器
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临床诊断
✓ 已验证
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Wentao Wang,Yingyu Zhang,Fei Zhang,Jing Gao,Zhidan Yu,Lifeng Li,Xianwei Zhang,Ligong Hou,Qiumei Zhou,Wancun Zhang
Sensitive, specific and simple quantitative and qualitative detection of nucleic acids is crucial for clinical diagnosis, especially in resource-limited settings. Therefore, in this study, an apurinic/apyrimidinic endonuclease 1 (APE1)-assisted cyclic amplification biosensor based on rolling circle amplification (RCA), referred to as A-RCA, was developed using molecular beacons as signal probes for highly sensitive and specific quantitative and qualitative detection of nucleic acids. The experimental results demonstrated that A-RCA exhibits exceptional sensitivity, with a detection limit of 19.5 aM and 14.3 aM for Mycoplasma pneumoniae ( M. pneumoniae ) DNA and M. pneumoniae RNA, respectively. In addition, A-RCA exhibits a broad linear range extending from 0.1 fM to 0.1 nM, and high specificity, enabling it to distinguish M. pneumoniae from other common respiratory pathogens. More importantly, a fluorescent test strip approach based on A-RCA (A-RCA-strip) was developed, enabling sensitive and specific point-of-care testing for the qualitative detection of nucleic acids. Similarly, the A-RCA-strip demonstrates high sensitivity, capable of detecting target signals as low as 0.1 fM, and exhibits high specificity in distinguishing M. pneumoniae from other common respiratory pathogenic microorganisms. Clinical detection showed that both A-RCA and A-RCA-strip could accurately identify M. pneumoniae , with 100 % concordance to established clinical methods. As a proof of concept, A-RCA and A-RCA-strip can detect other nucleic acid targets by simply changing the target sequence, providing a robust and versatile strategy for developing new clinical detection methods.
研究领域:
分子诊断
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临床检验医学
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分子生物学
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感染性疾病
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转化医学
✓ 已验证
DOI
高影响因子
BMC Veterinary Research
·
2022
·Vol. 18
·期 1
·1-14
·
IF 2.792
Hao Fei,Xie Xing,Feng Zhixin,Chen Rong,Wei Yanna,Liu Jin,Xiong Qiyan,Shao Guoqing,Lin Johnson
Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear.Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified, suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli, and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD+. Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains.Conclusion sNOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis.
研究领域:
微生物学
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兽医学
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分子生物学
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病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Tao Dong,Guangze Sun,Wanjian Liu,Yunze Long,Aihua Liu
Acute respiratory infections (ARIs) are among the leading causes of morbidity and mortality worldwide, with Mycoplasma pneumoniae (M. pneumoniae) and Chlamydia pneumoniae (C. pneumoniae) being major pathogens responsible for community-acquired respiratory infections. Rapid and accurate diagnosis is critical for improving patient outcomes and effectively controlling disease transmission. Herein, we report a novel dual-channel lateral flow immunoassay (DC-LFIA) platform based on the Co3O4@polydopamine (PDA) nanocomposite with outstanding extinction coefficient, oxidase-like activity, and magnetic properties for the simultaneous detection of specific IgM antibodies (Ab) against these two pathogens in human serum. The magnetic properties of the Co3O4@PDA nanocomposite enable rapid enrichment of trace target Abs from large-volume serum samples, with a high extinction coefficient for direct colorimetric visualization and an excellent oxidase-like catalytic activity for signal enhancement. On the basis of the multifunctional Co3O4@PDA nanocomposite, rapid dual-mode detection of serum Abs was achieved within 12 min. By the direct colorimetric mode, the visual limit of detection (vLOD) for M. pneumoniae Ab and C. pneumoniae Ab reached 2 and 1 ng/mL, respectively, while the oxidase-like catalyzing signal amplification mode further lowered the vLODs to 0.2 and 0.1 ng/mL, accordingly. Additionally, the signal normalization using a 3D-printed device significantly enhanced assay accuracy and reproducibility. When applied to 96 clinical serum samples, compared to commercial kits, this multifunctional Co3O4@PDA-based DC-LFIA demonstrated superior performance, providing a reliable approach for simultaneous antibody detection in respiratory infection diagnosis. Overall, this work contributes a valuable diagnostic tool to the field of rapid detection of ARIs.
研究领域:
免疫学
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临床检验医学
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纳米生物技术
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传染病学
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诊断学
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转化医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2024
·
IF 2.7
Huiqing Wang,Yang Li,Lin Tian,Xinyi Li,Qian Gao,Yaru Liu,Cuiping Ma,Qing Wang,Chao Shi
A concise and rapid detection method for Mycoplasma pneumoniae is urgently required due to its severe impact on human health. To meet such a need, this study proposed and constructed an innovative point-of-care testing (POCT) platform that consists of a hydrogen ion-selective loop-mediated isothermal amplification (H+-LAMP) sensor and an electrochemical detection device. The H+-LAMP sensor successfully integrated the working and reference electrodes and converted the H+ generated during the LAMP process into an electrochemical signal. High sensitivity and stability for pathogen detection were also achieved by treating the working electrode with an electrodeposited polyaniline solid contact layer and by using an ion-selective membrane. As a result, the sensor shows a sensitivity of 68.26 mV per pH, a response time of less than 2 s, and a potential drift of less than 5 mV within one hour, which well meets the urgent need. The results also demonstrated that the detection limit for Mycoplasma pneumoniae was lowered to 1 copy per μL, the nucleic acid extraction and detection process could be completed in 30 minutes, and the impact of interfering ions on the sensor was negligible. Validation with 20 clinical samples yielded satisfactory results. More importantly, the storage lifespan of such an electrochemical sensor is over seven days, which is a great advantage for on-site pathogen detection. Therefore, the hydrogen ion-selective sensor constructed in this investigation is particularly suitable as a core component for instant pathogen detection platforms.
研究领域:
生物医学工程
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微生物学
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电化学
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即时诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2022
·
IF 5.195
Zheng Liu,Wen-ting Dong,Wen-feng Wei,Jin-hai Huo,Wei-ming Wang
Ethnopharmacological relevance Traditional Chinese medicine categorizes Mycoplasma pneumoniae pneumonia as “lung heat”, and treatment with heat clear and detoxify. Traditional Chinese medicine believes that the lungs and intestines come from the same source, and the intestine is related to pneumonia. This is the same as the gut-lung axis theory. Qinbaiqingfei concentrate pills (QBs) were modified based on Cough San in the ancient medical book Medical Awareness . It clears lung heat, moisturizes the lungs and dredges collaterals, and has a good ability to treat Mycoplasma pneumoniae . Aim of the study A rat model of Mycoplasma pneumoniae was established. From the aspect of intestinal flora and mucosal immunity, the potential mechanism of the QBs was researched. Materials and methods First, the content of Mycoplasma pneumoniae in lung tissue and the levels of the inflammatory factors IL-4, IL-10, TNF-α and INF-γ were detected. To determine the expression of NF-kB related proteins in lung tissue, which can understand the ability in treating disease. Next, metagenomic sequencing was performed to detect changes in short-chain fatty acids, proving the ability of the drug to regulate intestinal microecology. Finally, HDAC , LPS , SIgA, etc. were detected to facilitate the correlation of the overall experimental indicators. Results QBs reduces the levels of IL-4, IL-10, TNF-α and INF-γ in the serum by inhibiting the expression of MyD88 , IKKα, IκBα, and NF-κB p65 in lung tissue. In addition, QBs restores the ratio of gram-negative bacteria to gram-positive bacteria in the intestine, restores the secretion of acetic acid, propionic acid , butyric acid , isobutyric acid and isovaleric acid , and promotes the secretion of NF-κB p65 and SIgA by HDAC1/3. The result is that the lung tissue is repaired and the proliferation of Mycoplasma pneumoniae is inhibited. Conclusions From the “gut-lung axis”, a new research perspective was discovered. QBs intervened in the intestines and lungs to treat Mycoplasma pneumoniae .
研究领域:
民族药理学
·
微生物学
·
免疫学
·
中医药
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2022
·
IF 3.571
Yake Duan,Xin Zhang,Yong Li,Xiaoli Zhao,Xiaowen Zhao,Lei Chen,Chao Shi,Cuiping Ma,Xiujuan Wang
Herein, we developed a facile integrated Mycoplasma pneumoniae diagnosis platform by combining amino-modified silica membrane (AMSM)-based nucleic acids fast extraction and enrichment with colorimetric isothermal amplification detection. AMSM demonstrates a strong ability to capture and enrich nucleic acids in complicated biological matrices, and the purified AMSM/nucleic acids composite could be directly used to perform isothermal amplification including denaturation bubble-mediated strand exchange amplification (SEA) and loop-mediated isothermal amplification (LAMP) reactions. Through comparing clinical specimens, excellent performance of AMSM-based SEA assay with 93.33% sensitivity and 100% specificity relative to real-time PCR was observed, and for AMSM-based LAMP was 96.67% and 100%, respectively. The diagnostic procedure could be completed within 55 min, and the colorimetric-based visual result further alleviates the use of sophisticated equipment. The proposed approach possesses great potential as a simple and time-saving alternative for point-of-care testing (POCT) of M. pneumoniae in resource-limited regions.
研究领域:
生物技术
·
分子生物学
·
临床诊断
·
微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
·
2021
·Vol. 36
·期 5
·e9234
·
IF 2.419
Jing Li,Yunhua Fu,Wei Jing,Jie Li,Xin Wang,Jialing Chen,Shuxin Sun,Hao Yue,Yulin Dai
Rationale Mycoplasma pneumoniae has become one of the common pathogens causing pediatric respiratory infections. In clinical diagnosis, throat swabs are very difficult to obtain from children, and there is a possibility of false positive results; hence, there are few clinically available diagnostic methods. Methods In this study, Q Exactive liquid chromatography/tandem mass spectrometry was used to analyze the metabolites in the urine of healthy children (HC) and M. pneumoniae pneumonia in children (MPPC) patients. A multivariate statistical analysis was performed to screen the differential metabolites. Based on the HMDB and KEGG, the possible metabolic pathways subject to biological alteration were identified. Results Compared with HC, 73 different metabolites in MPPC patients disrupted nine metabolic pathways through different change trends; after integrating various parameters, 20 significantly different metabolites were identified as MPPC potential biomarkers. Through the above two analysis modes, acetylphosphate and 2,5-dioxopentanoate were both screened out and used as potential biomarkers for the early diagnosis of MPPC for the first time. Conclusions The characterization of 20 potential biomarkers provides a scientific basis for predicting and diagnosing MPPC. This article further indicates that urine metabolic profiling has great potential in diagnosing MPPC and can effectively prevent the disease from causing further deterioration.
研究领域:
儿科
·
传染病学
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代谢组学
·
质谱分析
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2021
·
IF 5.64
Xie Xing,Hao Fei,Chen Rong,Wang Jingjing,Wei Yanna,Liu Jin,Wang Haiyan,Zhang Zhenzhen,Bai Yun,Shao Guoqing,Xiong Qiyan,Feng Zhixin
Mycoplasma hyopneumoniae (Mhp) is the main pathogen that causes enzootic pneumonia, a disease that has a significant impact on the pig industry worldwide. The pathogenesis of enzootic pneumonia, especially possible virulence factors of Mhp, has still not been fully elucidated. The transcriptomic and proteomic analyses of different Mhp strains reported in the literature have revealed differences in virulence, and differences in RNA transcription levels between high- and low-virulence strains initially indicated that nicotinamide adenine dinucleotide (NADH)-dependent flavin oxidoreductase (NFOR) was related to Mhp pathogenicity. Prokaryotic expression and purification of the NFOR protein from Mhp were performed, a rabbit-derived polyclonal antibody against NFOR was prepared, and multiple sequence alignment and evolutionary analyses of Mhp NFOR were performed. For the first time, it was found that the NFOR protein was conserved among all Mhp strains, and NFOR was localized to the cell surface and could adhere to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Adhesion to hTERT-PBECs could be specifically inhibited by an anti-NFOR polyclonal antibody, and the rates of adhesion to both high- and low-virulence strains, 168 and 168L, significantly decreased by more than 40%. Moreover, Mhp NFOR not only recognized and interacted with host fibronectin and plasminogen but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by hTERT-PBECs incubated with Mhp NFOR was significantly positively correlated with the virulence of Mhp. Overall, in addition to being a metabolic enzyme related to oxidative stress, NFOR may also function as a potential novel virulence factor of Mhp, thus contributing to the pathogenesis of Mhp; these findings provide new ideas and theoretical support for studying the pathogenic mechanisms of other mycoplasmas.
研究领域:
微生物学
·
病理学
·
兽医学
·
分子生物学
·
细胞生物学
✓ 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）