1.Epidemic characteristics and disease burden of brucellosis in Tongliao City in 2018 - 2023
Shanhong LIU ; Tu BA ; Li MA ; Na GUAN ; Lin XI ; Na TA
Journal of Public Health and Preventive Medicine 2026;37(2):74-78
Objective To comprehensively analyze the current epidemic characteristics and disease burden of brucellosis in Tongliao City, and to provide a basis for the prevention and control strategy of brucellosis in Tongliao City. Methods The report data of brucellosis in Tongliao City from 2018 to 2023 were collected. Descriptive methods were used for data analysis, and the disability-adjusted life years and indirect economic losses were calculated. Results From 2018 to 2023, a total of 22 034 cases were reported in Tongliao City, with an average annual incidence of 136.17/100 000. The incidence was statistically different between men and women ( χ2=12.23, P=0.032). The majority of cases were farmers (94.25%), followed by herdsmen (1.67%). The age group was concentrated between 30-60 years old (79.30%), among which the majority of cases were in the 40-50 years group (6 883/22 034). The onset time had seasonal characteristics, and the peak period was from March to August (the seasonal index was between 115.40%-151.29%). In terms of regional distribution, cases were reported in all counties (banners). The average annual incidence was highest in Kulun Banner (233.85/100 000) and Zalut Banner (210.13/100 000), and lowest in Keerqin District (42.28/100 000) and Holingol City (31.87/100 000). The analysis of disease burden showed that a total of 677.55 person-years (YLD) were lost from 2018 to 2023, with an average annual loss of 112.92 person-years. The total indirect economic loss was 59.3576 million yuan, with an average annual loss of 9.892 9 million yuan, and the people over 60 years old had the lowest annual loss. Conclusion The overall brucellosis epidemic in Tongliao City has shown a fluctuating downward trend. The epidemic prevention and control should be strengthened in farmers, people aged 40-50 years old, and areas such as Zalut Banner and Kulun Banner to further control the epidemic of brucellosis.
2.Analysis of Variety Characteristics and Patterns of Marketed Traditional Chinese Patent Medicines for Treating Chronic Gastritis
Daiyue DING ; Changyue SONG ; Shuangfei DENG ; Siyu LI ; Xiangying KONG ; Xiaohui SU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):252-260
ObjectiveThis study aims to systematically review the marketed traditional Chinese patent medicines for treating chronic gastritis (CG) in China. By analyzing their variety characteristics and prescription patterns, it seeks to provide references for clinical syndrome differentiation-based drug selection, treatment method optimization, and the design of high-quality clinical research. MethodsInformation on marketed traditional Chinese patent medicines for treating CG was collected. Microsoft Excel software was used to collate and statistically analyze representative drugs for each pathological stage, market status, syndrome types, and other contents. The Ancient and Modern Medical Case Cloud Platform (V2.3.9) was employed to analyze the formula composition patterns of standardized prescriptions. ResultsA total of 141 marketed traditional Chinese patent medicines for treating CG in China were included. Based on the disease's pathological progression, they can be classified into drugs for non-atrophic gastritis, atrophic gastritis, and precancerous lesions. Post-marketing research reveals that relevant evaluation is only conducted on 17 drugs, of which 2 involve pharmacoeconomic studies and 14 possess standardized evidence-based evidence. The primary dosage forms were capsules, granules, and tablets. From the 100 prescriptions screened according to inclusion/exclusion criteria, the varieties indicated for the stomach collateral stasis syndrome in atrophic gastritis accounted for the highest proportion. The main efficacy distributions were clearing heat, detoxifying, and relieving pain by promoting Qi circulation. Core drugs included Glycyrrhizae Radix et Rhizome, Paeoniae Radix Alba, and Aucklandiae Radix. Medicinal properties were predominantly warm and neutral. Flavors were mainly bitter, pungent, and sweet. The drugs primarily entered the spleen and stomach meridians. Analysis of the package inserts reveals that 67 products list "contraindications", 110 include "precautions", and 23 explicitly state "adverse reactions". ConclusionTraditional Chinese patent medicines for treating CG hold unique value in clinical practice. However, currently there are challenges such as insufficient clarity in syndrome type descriptions within package inserts and a relative lack of high-level evidence-based medical evidence, as well as pharmacoeconomic evaluations. Future efforts should focus on addressing these shortcomings by advancing research on syndrome characteristics and medication patterns based on syndrome differentiation, systematically conducting pharmacoeconomic evaluations, strengthening the accumulation of high-level evidence-based evidence, and, on this basis, improving patient medication adherence. This will comprehensively enhance the clinical application value and scientific connotation of this category of drugs.
3.Material Basis of Entada phaseoloides Against Neuropathic Pain Based on in vitro Chemical Profiling and in vivo Absorbed Constituents
Sensen LI ; Hanxue WU ; Yuanbin LI ; Limin ZHAO ; Xiaohui SU ; Hui XIONG ; Weihua MA ; Chao WANG ; Zhaochen MA ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(18):88-97
ObjectiveTo systematically characterize the in vitro chemical constituents of Entada phaseoloides and its absorbed prototype compounds and metabolites in a neuropathic pain (NP) model, thereby elucidating the potential pharmacological material basis underlying its analgesic activity. MethodsUltra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to analyze the constituents of the 80% methanol extract of E. phaseoloides. Thirty-six mice were divided into a blank group, low-, medium-, and high-dose E. phaseoloides extract groups (25, 50, and 100 mg·kg-1, respectively), an aspirin group (400 mg·kg-1), and a morphine group (10 mg·kg-1), with six mice in each group. The tail-flick threshold was measured. Twenty-four mice were randomly divided into a blank group and low-, medium-, and high-dose E. phaseoloides extract groups (25, 50, and 100 mg·kg-1, respectively), with six mice in each group, and motor coordination was evaluated. A spinal nerve ligation (SNL) model was established in mice. Twenty-four mice were randomly divided into a sham-operated (Sham) group, an SNL group, an SNL + E. phaseoloides extract group (750 mg·kg-1), and an SNL + pregabalin group (20 mg·kg-1), with 6 mice in each group. Mechanical allodynia and thermal hyperalgesia were evaluated. Behavioral assessments were combined to determine the optimal onset time of the analgesic effect, and plasma samples were collected at this time point. Absorbed components and metabolites were systematically identified based on accurate mass, MS/MS fragmentation patterns, retention time, and comparisons with literature data and reference standards. Meanwhile, network pharmacology, target similarity analysis, and molecular docking validation were integrated to evaluate the potential effects of absorbed components on disease intervention. ResultsCompared with the blank group, the high-dose E. phaseoloides extract group and morphine group showed significantly increased pain response thresholds (P<0.01), without affecting motor coordination in normal mice. Compared with the Sham group, the SNL group exhibited significantly decreased thresholds for mechanical allodynia and thermal hyperalgesia (P<0.01). Compared with the SNL group, the SNL + E. phaseoloides extract group and SNL + pregabalin group showed significantly increased thresholds for mechanical allodynia and thermal hyperalgesia (P<0.01). The analgesic effect of E. phaseoloides was most significant at 2 h after administration. A total of 70 constituents were identified in vitro, and 22 prototype compounds and 29 metabolites were detected in plasma. The targets of the absorbed components were mainly enriched in biological processes associated with pain signal perception and transmission and showed a high degree of overlap with disease genes related to neuropathic pain. In addition, multiple components exhibited strong binding energies with core pain-related targets. Based on integrated analysis of in vitro content, plasma exposure levels, and target associations, ketanthamide A-β-D-pyranoside and ketengzide were identified as the most representative pharmacologically active substances responsible for the analgesic effect. ConclusionThis study systematically elucidates the material basis of E. phaseoloides intervention in neuropathic pain from three aspects, including chemical constituent characteristics, in vivo behavioral characteristics, and potential targets, providing important theoretical foundations and experimental support for further studies on its pharmacological mechanism and analgesic drug development.
4.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
5.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
6.Characteristic Analysis of Effective Components and Compounds of TCM for Prevention and Treatment of Breast Cancer Based on Wnt/β-catenin Signaling Pathway Targeting
Haoyang WANG ; Lin GUO ; Hui ZHAO ; Lihua CAO ; Na LI ; Mingsan MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):282-290
Breast cancer is a kind of malignant tumor with a complex mechanism, and its morbidity and mortality are increasing year by year, which seriously threatens women's health. At present, the main clinical treatments are surgical resection, radiotherapy, chemotherapy, and drug therapy, but they are often accompanied by side effects and adverse reactions, which affect the therapeutic effect. Traditional Chinese medicine (TCM) has the advantages of multi-component and multi-target treatment in the fight against breast cancer. The wnt/β-catenin signaling pathway is one of the classic pathways in cancer research. Abnormally activated Wnt/β-catenin signaling pathway inhibits β-catenin degradation by blocking the formation of Axin/glycogen synthase kinase 3β/adenomatous polyposis coli complex, thus promoting β-catenin nuclear metastasis, and it binds to T cell transcription factor/lymphoenhancer factor-1 to initiate downstream target genes and further interfere with the proliferation, migration, and invasion of tumor cells to affect the tumor process. Previous studies have shown that TCM monomers and compounds can mediate the Wnt/β-catenin signaling pathway to inhibit the malignant phenotype of breast cancer cells, thus playing an anti-breast cancer role, and the biochemical process involved in the regulation of therapeutic drugs has not been systematically combed. By analyzing and collating Chinese and foreign literature at the present stage, this paper discussed the association mechanism between Wnt/β-catenin signaling pathway and breast cancer and analyzed the internal mechanism of TCM monomers and compounds in mediating Wnt/β-catenin signaling pathway to exert anti-breast cancer effect. The statistical results showed that the flavonoids, alkaloids, and terpenoids in TCM monomers could target the Wnt/β-catenin signaling pathway and block the further development of malignant phenotype of breast cancer cells. TCM compounds with functions of clearing heat and detoxifying, promoting blood circulation and removing blood stasis, and tonifying kidney and liver were commonly used to intervene in the Wnt/β-catenin signaling pathway to prevent breast cancer. Compared with the current inhibitors of Wnt/β-catenin signaling pathway, the application of TCM monomers and compounds is expected to bring low-toxicity and high-efficiency breast cancer treatment drugs to the clinical practice, and the existing results provide a reference for the subsequent screening, research, and development of TCM small-molecule compounds and TCM compounds against breast cancer.
7.Interventional Effect and Mechanisms of Renqing Mangjue on MNNG-induced Malignant Transformation of Gastric Mucosal Epithelial Cells
Peiping CHEN ; Fengyu HUANG ; Xinzhuo ZHANG ; Xiangying KONG ; Ziqing XIAO ; Yanxi LI ; Xiaohui SU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):69-77
ObjectiveThis study aimed to investigate the intervention effect of Renqing Mangjue on the malignant transformation of gastric mucosal epithelial cells induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and to explore its molecular mechanism in preventing precancerous lesions of gastric cancer based on the cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodsHuman gastric mucosal epithelial cells (GES-1) were initially induced by MNNG to establish a precancerous cell model (MC cells). The effective concentration of MNNG for inducing malignant transformation in GES-1 cells was screened using the cell proliferation activity decection (CCK-8) assay, and the effective concentration of Renqing Mangjue for inhibiting the proliferation of transformed GES-1 cells was also determined. GES-1 cells were divided into a blank control group, a model group, and treatment groups with Renqing Mangjue at concentrations of 1, 3, 10, and 30 mg·L-1. Furthermore, the effects of Renqing Mangjue on the migratory ability and epithelial-mesenchymal transition (EMT) characteristics of GES-1 malignant transformed cells were evaluated using Transwell migration assays, wound healing assays, and real-time quantitative reverse transcription polymerase chain reaction (Real-time PCR). Additionally, candidate chemical components and target sites of Renqing Mangjue were obtained from the TCMIP v2.0 database, and disease targets at various stages of gastric cancer precursors were sourced from the Gene Expression Omnibus (GEO) database. Pathway enrichment analysis was performed using the Metascape database to predict the potential mechanisms of action of Renqing Mangjue. Finally, the protective mechanism of Renqing Mangjue against gastric cancer precursors was validated through Western blot analysis. ResultsAt a concentration of 20 μmol·L-1, MNNG exhibited an inhibition rate of approximately 50% on GES-1 cells (P<0.01), and at this concentration, the GES-1 cells displayed biological characteristics indicative of malignant transformation. In contrast, Renqing Mangjue had no significant effect on the proliferation of normal GES-1 cells, but significantly inhibited the proliferation of MC cells (P<0.01) and markedly reduced their migratory capacity (P<0.01). Moreover, it also increased the mRNA expression level of E-cadherin during the EMT process (P<0.05), while inhibiting the expression of both N-cadherin and the transcription factor Snail mRNA (P<0.05, P<0.01). Network predictions suggested that Renqing Mangjue may prevent gastric cancer precursors through modulating the cGMP/PKG and MAPK/ERK signaling pathways. Furthermore, Western blot results indicated that Renqing Mangjue upregulated the expression of PKG and NPRB (B-type natriuretic peptide receptor) proteins in the cGMP/PKG pathway (P<0.01), while downregulating the expression of the downstream proteins MEK and ERK (P<0.05, P<0.01). ConclusionIn summary, Renqing Mangjue can prevent gastric cancer precursors by inhibiting the proliferation and migration of malignant transformed GES-1 cells, thereby delaying the EMT process. The underlying mechanisms may be related to the activation of the cGMP/PKG pathway and the inhibition of the MEK/ERK signaling pathway.
8.Investigation on the mechanisms of Colquhounia Root Tablets in reversing vascular endothelial cell dysfunction of rheumatoid arthritis via modulating NOD2/SMAD3/VEGFA signaling axis
Bing-bing CAI ; Ya-wen CHEN ; Tao LI ; Yuan ZENG ; Yan-qiong ZHANG ; Na LIN ; Xia MAO ; Ya LIN
Acta Pharmaceutica Sinica 2025;60(2):397-407
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation, joint destruction, and functional impairment. Angiogenesis plays a key role in the pathological progression of RA with dysfunction of endothelial cells to promote synovial inflammation, sustain pannus formation, subsequently leading to joint damage. Colquhounia Root Tablets (CRT), a Chinese patent drug, has shown a satisfying clinical efficacy in treating RA, while the underlying mechanism by which CRT inhibits RA-associated angiogenesis remains unclear. In this study, we applied a research approach combining transcriptomic data analysis, bio-network mapping, and
9.Survey of influenza awareness of community residents in nanchang during routine epidemic prevention and control
Shenggao LI ; Huai JIANG ; Yao ZHU ; Kerong FANG ; Xinya LIN ; Qingfeng ZHU ; Na TIAN
Modern Hospital 2025;25(5):787-789
Objective To investigate influenza awareness of community residents in Nanchang under routine epidemic management using the Knowledge-Attitude-Practice(KAP)framework.Methods A structured questionnaire,including four do-mains(disease overview,prevention,diagnosis,and treatment),was designed and administered via street and online surveys from December 2023 to February 2024.Statistical analyses,including chi-square tests and logistic regression,were performed to identify determinant factors.Results A total of 303 valid questionnaires were retrieved,with an effective rate of approximately 94.7%.Community residents in Nanchang demonstrated a certain degree of understanding of the overview,prevention,diagno-sis,and treatment of influenza,although some gaps were identified.Univariate analysis revealed statistically significant differ-ences in influenza cognition scores among urban and rural residents(P<0.05),residents with different educational levels(P<0.05)and exposure levels to influenza education(P<0.05).Multivariate regression analysis identified these as influencing fac-tors for influenza awareness.Conclusion Collaborative efforts from the government,individuals,and various societal sectors are essential to enhance the public's overall ability to prevent and control influenza.
10.Establishment and application of ultra-fast real-time PCR for Brucella detection
Zhen-na XU ; Zhi-peng WU ; Wei-bin HONG ; Zhi-shen GUAN ; Qi-ming LIN ; Zuan-lan MO ; Yi-fei YE ; Hai-yan XIE ; Min LI ; Yan-qiu ZHU ; Xiao-jun LI ; Xian-peng ZHANG
Chinese Journal of Zoonoses 2025;41(3):278-283
This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20%to 0.91%,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100%compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.


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