1.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
2.Systematic review of the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures
Feifei HAN ; Jing TIAN ; Lingyan QIAO ; Haili YIN ; Xing WEI ; Lili FENG
Chinese Journal of Trauma 2025;41(7):675-681
Objective:To systematically review the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures.Methods:PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang Database and VIP Database were systematically searched to collect literature on the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures from inception to June 30, 2024. The languages were limited to Chinese and English. Two researchers screened the literature according to the inclusion and exclusion criteria. Data extraction was performed using the checklist for critical appraisal and data extraction for systematic reviews of prediction modeling studies (CHARMS), encompassing basic study characteristics, model development features, and model performance metrics. The predictors, validation methods, presentation formats, and predictive performance of the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures were evaluated. The prediction model risk-of-bias assessment tool (PROBAST) was employed to assess risk of bias and applicability of the included studies.Results:A total of 11 studies, comprising of 16 prediction models, were included, with a total sample size of 283-1 508 patients and a pulmonary infection incidence rate of 5.4%-16.25%. The independent predictive factors repeatedly included in the models were age, American Society of Anesthesiologists (ASA) scale, preoperative comorbidities, chronic obstructive pulmonary disease (COPD), preoperative albumin level, white blood cell count (WBC), and C-reactive protein (CRP) level. The models were internally validated in 7 studies and externally validated in 3. The models were visualized in the form of a nomogram in 7 studies and a web-based risk calculator in 1. Model prediction performance was analyzed: (1) In terms of the discrimination, 9 studies reported the area under the receiver operating characteristic curve (AUC), with the overall AUC range of 0.664-0.905. (2) In terms of the calibration, 5 studies had Hosmer-Lemeshow test, with the P-values all above 0.05; 2 studies reported the calibration plots, with the slopes close to 1 and the Brier scores of 0.016 and 0.112; 4 studies reported the sensitivity of the models of 73.91%-92.40% and specificity of 57.10%-92.41%. According to PROBAST, all 11 studies exhibited certain risk of bias while maintaining favorable applicability. Conclusions:Age, ASA scale, preoperative comorbidities, COPD, preoperative albumin level, WBC, and CRP level are found to be independent predictive factors repeatedly reported in the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures. The existing models demonstrate a robust overall prediction performance despite certain risks of bias.
3.Diagnosis, treatment, and genetic analysis of five cases of primary atypical hemolytic uremic syndrome
Weiying HE ; Fen TIAN ; Jing LI ; Runhong HAN ; Guangqun XING
Chinese Journal of Internal Medicine 2025;64(6):549-557
A retrospective analysis was conducted on the clinical characteristics, renal pathology, genetic testing, and treatment of five patients —two males and three females—diagnosed with primary atypical hemolytic uremic syndrome (aHUS) in the Department of Nephrology at the Affiliated Hospital of Qingdao University from February 2022 to June 2024. The patients′ ages at disease onset ranged from 14 to 29 years. Four patients experienced prodromal infection symptoms. At disease onset, serum creatinine levels ranged from 168.5 to 1 230.2 μmol/L. All patients presented with hematuria, proteinuria, hypertension, non-immune hemolytic anemia, thrombocytopenia, elevated lactate dehydrogenase (LDH), and fragmented red blood cells in peripheral blood (0.5%-6.0%). Serum haptoglobin levels were below the normal lower limit in all cases. Four patients demonstrated decreased serum complement C3, while one maintained normal serum complement C3 throughout the course of the disease. One patient exhibited serum factor H concentrations below the normal lower limit. Another patient tested positive for anti-factor H antibodies. Renal biopsies were performed on four patients. Electron microscopy revealed typical acute-phase pathological features of aHUS in three cases, including glomerular endothelial cell swelling and widened subendothelial spaces. One patient demonstrated ischemic and atrophic changes in the glomerular capillaries, while another had concurrent membranous nephropathy. Whole-exome high-throughput sequencing related to aHUS was performed in all five patients, revealing heterozygous gene mutations in each case. Complement-related gene mutations, typically occurring in a heterozygous state, are prevalent in aHUS patients. The eight heterozygous gene variations identified in this study were absent from existing databases of known aHUS-associated pathogenic mutations. Four patients received eculizumab treatment at varying time points following diagnosis, resulting in differing clinical outcomes. The patient positive for anti-factor H antibodies was treated with rituximab. The patient with membranous nephropathy initiated combination therapy with rituximab and eculizumab after six months of eculizumab monotherapy. Following treatment, all five patients achieved complete cessation of intravascular mechanical hemolysis, with normalization of LDH and platelet levels, as well as varying degrees of renal function recovery. From a pathophysiological perspective, the timely administration of the complement C5 inhibitor eculizumab can rapidly induce clinical remission, reduce the incidence of end-stage renal disease, and improve prognosis in patients with aHUS.
4.Systematic review of the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures
Feifei HAN ; Jing TIAN ; Lingyan QIAO ; Haili YIN ; Xing WEI ; Lili FENG
Chinese Journal of Trauma 2025;41(7):675-681
Objective:To systematically review the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures.Methods:PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang Database and VIP Database were systematically searched to collect literature on the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures from inception to June 30, 2024. The languages were limited to Chinese and English. Two researchers screened the literature according to the inclusion and exclusion criteria. Data extraction was performed using the checklist for critical appraisal and data extraction for systematic reviews of prediction modeling studies (CHARMS), encompassing basic study characteristics, model development features, and model performance metrics. The predictors, validation methods, presentation formats, and predictive performance of the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures were evaluated. The prediction model risk-of-bias assessment tool (PROBAST) was employed to assess risk of bias and applicability of the included studies.Results:A total of 11 studies, comprising of 16 prediction models, were included, with a total sample size of 283-1 508 patients and a pulmonary infection incidence rate of 5.4%-16.25%. The independent predictive factors repeatedly included in the models were age, American Society of Anesthesiologists (ASA) scale, preoperative comorbidities, chronic obstructive pulmonary disease (COPD), preoperative albumin level, white blood cell count (WBC), and C-reactive protein (CRP) level. The models were internally validated in 7 studies and externally validated in 3. The models were visualized in the form of a nomogram in 7 studies and a web-based risk calculator in 1. Model prediction performance was analyzed: (1) In terms of the discrimination, 9 studies reported the area under the receiver operating characteristic curve (AUC), with the overall AUC range of 0.664-0.905. (2) In terms of the calibration, 5 studies had Hosmer-Lemeshow test, with the P-values all above 0.05; 2 studies reported the calibration plots, with the slopes close to 1 and the Brier scores of 0.016 and 0.112; 4 studies reported the sensitivity of the models of 73.91%-92.40% and specificity of 57.10%-92.41%. According to PROBAST, all 11 studies exhibited certain risk of bias while maintaining favorable applicability. Conclusions:Age, ASA scale, preoperative comorbidities, COPD, preoperative albumin level, WBC, and CRP level are found to be independent predictive factors repeatedly reported in the risk prediction models for postoperative pulmonary infection in elderly patients with hip fractures. The existing models demonstrate a robust overall prediction performance despite certain risks of bias.
5.Diagnosis, treatment, and genetic analysis of five cases of primary atypical hemolytic uremic syndrome
Weiying HE ; Fen TIAN ; Jing LI ; Runhong HAN ; Guangqun XING
Chinese Journal of Internal Medicine 2025;64(6):549-557
A retrospective analysis was conducted on the clinical characteristics, renal pathology, genetic testing, and treatment of five patients —two males and three females—diagnosed with primary atypical hemolytic uremic syndrome (aHUS) in the Department of Nephrology at the Affiliated Hospital of Qingdao University from February 2022 to June 2024. The patients′ ages at disease onset ranged from 14 to 29 years. Four patients experienced prodromal infection symptoms. At disease onset, serum creatinine levels ranged from 168.5 to 1 230.2 μmol/L. All patients presented with hematuria, proteinuria, hypertension, non-immune hemolytic anemia, thrombocytopenia, elevated lactate dehydrogenase (LDH), and fragmented red blood cells in peripheral blood (0.5%-6.0%). Serum haptoglobin levels were below the normal lower limit in all cases. Four patients demonstrated decreased serum complement C3, while one maintained normal serum complement C3 throughout the course of the disease. One patient exhibited serum factor H concentrations below the normal lower limit. Another patient tested positive for anti-factor H antibodies. Renal biopsies were performed on four patients. Electron microscopy revealed typical acute-phase pathological features of aHUS in three cases, including glomerular endothelial cell swelling and widened subendothelial spaces. One patient demonstrated ischemic and atrophic changes in the glomerular capillaries, while another had concurrent membranous nephropathy. Whole-exome high-throughput sequencing related to aHUS was performed in all five patients, revealing heterozygous gene mutations in each case. Complement-related gene mutations, typically occurring in a heterozygous state, are prevalent in aHUS patients. The eight heterozygous gene variations identified in this study were absent from existing databases of known aHUS-associated pathogenic mutations. Four patients received eculizumab treatment at varying time points following diagnosis, resulting in differing clinical outcomes. The patient positive for anti-factor H antibodies was treated with rituximab. The patient with membranous nephropathy initiated combination therapy with rituximab and eculizumab after six months of eculizumab monotherapy. Following treatment, all five patients achieved complete cessation of intravascular mechanical hemolysis, with normalization of LDH and platelet levels, as well as varying degrees of renal function recovery. From a pathophysiological perspective, the timely administration of the complement C5 inhibitor eculizumab can rapidly induce clinical remission, reduce the incidence of end-stage renal disease, and improve prognosis in patients with aHUS.
6.Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients (version 2024)
Yao LU ; Yang LI ; Leiying ZHANG ; Hao TANG ; Huidan JING ; Yaoli WANG ; Xiangzhi JIA ; Li BA ; Maohong BIAN ; Dan CAI ; Hui CAI ; Xiaohong CAI ; Zhanshan ZHA ; Bingyu CHEN ; Daqing CHEN ; Feng CHEN ; Guoan CHEN ; Haiming CHEN ; Jing CHEN ; Min CHEN ; Qing CHEN ; Shu CHEN ; Xi CHEN ; Jinfeng CHENG ; Xiaoling CHU ; Hongwang CUI ; Xin CUI ; Zhen DA ; Ying DAI ; Surong DENG ; Weiqun DONG ; Weimin FAN ; Ke FENG ; Danhui FU ; Yongshui FU ; Qi FU ; Xuemei FU ; Jia GAN ; Xinyu GAN ; Wei GAO ; Huaizheng GONG ; Rong GUI ; Geng GUO ; Ning HAN ; Yiwen HAO ; Wubing HE ; Qiang HONG ; Ruiqin HOU ; Wei HOU ; Jie HU ; Peiyang HU ; Xi HU ; Xiaoyu HU ; Guangbin HUANG ; Jie HUANG ; Xiangyan HUANG ; Yuanshuai HUANG ; Shouyong HUN ; Xuebing JIANG ; Ping JIN ; Dong LAI ; Aiping LE ; Hongmei LI ; Bijuan LI ; Cuiying LI ; Daihong LI ; Haihong LI ; He LI ; Hui LI ; Jianping LI ; Ning LI ; Xiying LI ; Xiangmin LI ; Xiaofei LI ; Xiaojuan LI ; Zhiqiang LI ; Zhongjun LI ; Zunyan LI ; Huaqin LIANG ; Xiaohua LIANG ; Dongfa LIAO ; Qun LIAO ; Yan LIAO ; Jiajin LIN ; Chunxia LIU ; Fenghua LIU ; Peixian LIU ; Tiemei LIU ; Xiaoxin LIU ; Zhiwei LIU ; Zhongdi LIU ; Hua LU ; Jianfeng LUAN ; Jianjun LUO ; Qun LUO ; Dingfeng LYU ; Qi LYU ; Xianping LYU ; Aijun MA ; Liqiang MA ; Shuxuan MA ; Xainjun MA ; Xiaogang MA ; Xiaoli MA ; Guoqing MAO ; Shijie MU ; Shaolin NIE ; Shujuan OUYANG ; Xilin OUYANG ; Chunqiu PAN ; Jian PAN ; Xiaohua PAN ; Lei PENG ; Tao PENG ; Baohua QIAN ; Shu QIAO ; Li QIN ; Ying REN ; Zhaoqi REN ; Ruiming RONG ; Changshan SU ; Mingwei SUN ; Wenwu SUN ; Zhenwei SUN ; Haiping TANG ; Xiaofeng TANG ; Changjiu TANG ; Cuihua TAO ; Zhibin TIAN ; Juan WANG ; Baoyan WANG ; Chunyan WANG ; Gefei WANG ; Haiyan WANG ; Hongjie WANG ; Peng WANG ; Pengli WANG ; Qiushi WANG ; Xiaoning WANG ; Xinhua WANG ; Xuefeng WANG ; Yong WANG ; Yongjun WANG ; Yuanjie WANG ; Zhihua WANG ; Shaojun WEI ; Yaming WEI ; Jianbo WEN ; Jun WEN ; Jiang WU ; Jufeng WU ; Aijun XIA ; Fei XIA ; Rong XIA ; Jue XIE ; Yanchao XING ; Yan XIONG ; Feng XU ; Yongzhu XU ; Yongan XU ; Yonghe YAN ; Beizhan YAN ; Jiang YANG ; Jiangcun YANG ; Jun YANG ; Xinwen YANG ; Yongyi YANG ; Chunyan YAO ; Mingliang YE ; Changlin YIN ; Ming YIN ; Wen YIN ; Lianling YU ; Shuhong YU ; Zebo YU ; Yigang YU ; Anyong YU ; Hong YUAN ; Yi YUAN ; Chan ZHANG ; Jinjun ZHANG ; Jun ZHANG ; Kai ZHANG ; Leibing ZHANG ; Quan ZHANG ; Rongjiang ZHANG ; Sanming ZHANG ; Shengji ZHANG ; Shuo ZHANG ; Wei ZHANG ; Weidong ZHANG ; Xi ZHANG ; Xingwen ZHANG ; Guixi ZHANG ; Xiaojun ZHANG ; Guoqing ZHAO ; Jianpeng ZHAO ; Shuming ZHAO ; Beibei ZHENG ; Shangen ZHENG ; Huayou ZHOU ; Jicheng ZHOU ; Lihong ZHOU ; Mou ZHOU ; Xiaoyu ZHOU ; Xuelian ZHOU ; Yuan ZHOU ; Zheng ZHOU ; Zuhuang ZHOU ; Haiyan ZHU ; Peiyuan ZHU ; Changju ZHU ; Lili ZHU ; Zhengguo WANG ; Jianxin JIANG ; Deqing WANG ; Jiongcai LAN ; Quanli WANG ; Yang YU ; Lianyang ZHANG ; Aiqing WEN
Chinese Journal of Trauma 2024;40(10):865-881
Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.
7.Linezolid-induced fatal lactic acidosiss:an adverse reaction easily confu-sed with septic shock
Ran ZHU ; Shao-Wan YANG ; Xing-Han TIAN ; Xiao-Li LI ; Yan LIU
Chinese Journal of Infection Control 2024;23(7):897-900
Clinical data of 3 patients with linezolid-induced lactic acidosis treated in 2 hospitals in Beijing and Shandong were retrospectively analyzed,and relevant literatures at home and abroad were retrieved.The pathogene-sis,risk factors,clinical manifestations,and treatment scheme were valuated.Three patients received linezolid anti-infection treatment due to their disease condition.At different phases of treatment,they developed lactic acidosis that was difficult to be explained with septic shock.After discontinuing linezolid and receiving bedside continuous veno-venous hemofiltration(CVVH)treatment,patient's blood lactate levels decreased significantly.It is clinically diagnosed linezolid-induced lactic acidosis finally,but the initial diagnosis was septic shock by all doctors.After ac-tive treatment,2 patients recovered and 1 patient died.Linezolid-induced lactic acidosis is easily to be misdiagnosed as septic shock.Clinicians should enhance their understanding and recognition of the early diagnosis of the adverse reactions,take effective measures,so as to improve patient prognosis.
8.Risk factors of gastrointestinal bleeding after type A aortic dissection
Shi-Si LI ; Chun-Shui LIANG ; Tian-Bo LI ; Yun ZHU ; Han-Ting LIU ; Xing-Lu WANG ; Si ZHANG ; Rui-Yan MA
Journal of Regional Anatomy and Operative Surgery 2024;33(6):497-500
Objective To analyze the risk factors of gastrointestinal bleeding in patients with type A aortic dissection(TAAD)after Sun's operation.Methods The clinical data of 87 patients who underwent TAAD Sun's operation in our hospital from March 2021 to June 2022 were retrospectively analyzed.They were divided into the bleeding group and the non-bleeding group according to whether there was gastrointestinal bleeding after operation.The clinical data of patients in the two groups was compared and analyzed.The binary Logistic regression analysis was used to analyze the risk factors of gastrointestinal bleeding.The clinical predictor of postoperative gastrointestinal bleeding was analyzed by receiver operating characteristic(ROC)curve.Results In this study,there were 40 cases of postoperative gastrointestinal bleeding(the bleeding group)and 47 cases of non-bleeding(the non-bleeding group).Compared with the non-bleeding group,the bleeding group had a shorter onset time,a higher proportion of patients with hypertension history,a higher preoperative creatinine abnormality rate,more intraoperative blood loss,longer postoperative mechanical ventilation time,higher postoperative infection rate,and higher poor prognosis rate,with statistically significant differences(P<0.05).There was no statistically significant difference in the gender,age,gastrointestinal diseases history,smoking history,preoperative platelets,preoperative international normalized ratio(INR),preoperative alanine aminotransferase(ALT),preoperative aspartate aminotransferase(AST),preoperative γ-glutamyl transpeptidase(GGT),preoperative dissection involving abdominal aorta,operation time,intraoperative cardiopulmonary bypass time,intraoperative circulatory arrest time,intraoperative aortic occlusion time or intraoperative blood transfusion rate.Logistic regression analysis showed that hypertension history(OR=2.468,95%CI:0.862 to 7.067,P=0.037),preoperative creatinine>105 μmol/L(OR=3.970,95%CI:1.352 to 11.659,P=0.011),long postoperative mechanical ventilation time(OR=1.015,95%CI:0.094 to 1.018,P=0.041)and postoperative infection(OR=3.435,95%CI:0.991 to 11.900,P=0.012)were the independent risk factors for postoperative gastrointestinal bleeding in TAAD patients.ROC curve showed that the postoperative mechanical ventilation time exceeding 64 hours were the clinical predictor of postoperative gastrointestinal bleeding in TAAD patients.Conclusion The prognosis of TAAD patients with postoperative gastrointestinal bleeding after Sun's operation is poor.Hypertension history,preoperative acute renal insufficiency,long postoperative mechanical ventilation time and postoperative infection are closely related to postoperative gastrointestinal bleeding in TAAD patients after operation,which should be paid more attention to,and corresponding evaluation,early identification and early intervention should be made to improve the prognosis of patients.
9.Minimally invasive treatment and surgical injury control strategies for elderly patients with acute incarcerated ingui-nal hernias
Zhou-Wei XU ; Bai-Cheng DING ; Kai-Qiang WANG ; Tian-Le ZHAO ; Xing-Han LI ; Xing-Yu WANG
Chinese Journal of Current Advances in General Surgery 2024;27(8):622-626
Objective:To explore the application value and damage control of minimally inva-sive techniques in the treatment of acute incarcerated inguinal hernias in the elderly.Methods:In this study,62 elderly patients with acute incarcerated inguinal hernias admitted to the department of emergency surgery at the First Affiliated Hospital of Anhui Medical University from June 2018 to June 2023 were selected as the research subjects.After obtaining informed consent from the pa-tient's family for both treatment modalities,they were randomly divided into open surgery group and laparoscopic surgery group.Differences in clinical efficacy,perioperative indicators,post-operative complications,and prognostic follow-up of the two groups of patients were observed.Seven cases of elderly patients aged above 80 had many underlying diseases and poor tolerance during surgery.After treatment of lesions in the hernia contents,only damage control surgery for hernia sac high ligation was performed.Results:In comparison to patients treated with laparo-scopic surgery,there were statistically significant differences(P<0.05)in the open surgery group in clinical efficacy(efficacy,ineffectiveness,and overall effectiveness),perioperative indicators(length of stay,recovery time of digestive tract function,and VAS pain score),post-operative complica-tions,and prognostic follow-up(local mass,chronic pain,and ratio of second-stage hernia sur-gery).Seven patients treated according to injury control strategies all recovered and discharged from hospital after surgery.Conclusion:Emergency laparoscopic surgery for detecting incarcer-ated inguinal hernias in the elderly is safe and feasible.At the same time,it is essential to correctly assess the patient's vital signs during surgery.If necessary,surgery should be simplified to provide opportunities for follow-up treatment.
10.Establishment of a Multiplex Detection Method for Common Bacteria in Blood Based on Human Mannan-Binding Lectin Protein-Conjugated Magnetic Bead Enrichment Combined with Recombinase-Aided PCR Technology
Jin Zi ZHAO ; Ping Xiao CHEN ; Wei Shao HUA ; Yu Feng LI ; Meng ZHAO ; Hao Chen XING ; Jie WANG ; Yu Feng TIAN ; Qing Rui ZHANG ; Na Xiao LYU ; Qiang Zhi HAN ; Xin Yu WANG ; Yi Hong LI ; Xin Xin SHEN ; Jun Xue MA ; Qing Yan TIE
Biomedical and Environmental Sciences 2024;37(4):387-398
Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.

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