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Development of a droplet digital polymerase chain reaction assay for the sensitive detection of total and integrated hiv-1 DNA

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Chinese Medical Journal 2024年第6期137卷 729-736页

作者： Lin Yuan Zhiying Liu Xin Zhang Feili Wei Shan Guo Na Guo Lifeng Liu Zhenglai Ma Yunxia Ji Rui Wang Xiaofan Lu Zhen Li Wei Xia Hao Wu Tong Zhang Bin Su Beijing Key Laboratory for hiv/AIDS Research Clinical and Research Center for Infectious DiseasesBeijing Youan HospitalCapital Medical UniversityBeijing 100069China Beijing Institute of Hepatology Beijing Youan HospitalCapital Medical UniversityBeijing 100069China

Background:Total human immunodeficiency virus(hiv)DNA and integrated hiv DNA are widely used markers of hiv persistence.Droplet digital polymerase chain reaction(ddPCR)can be used for absolute quantification without needing a standard curve.Here,we developed duplex ddPCR assays to detect and quantify total hiv DNA and integrated hiv DNA.Methods:The limit of detection,dynamic ranges,sensitivity,and reproducibility were evaluated by plasmid constructs containing both the hiv long terminal repeat(LTR)and human CD3 gene(for total hiv DNA)and ACH-2 cells(for integrated hiv DNA).Forty-two cases on stable suppressive antiretroviral therapy(ART)were assayed in total hiv DNA and integrated hiv DNA.Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive(CD4^(+))T-cell counts,CD8^(+)T-cell counts and CD4/CD8 T-cell ratio,respectively.The assay linear dynamic range and lower limit of detection(LLOD)were also assessed.Results:The assay could detect the presence of hiv-1 copies 100%at concentrations of 6.3 copies/reaction,and the estimated LLOD of the ddPCR assay was 4.4 hiv DNA copies/reaction(95%confidence intervals[CI]:3.6-6.5 copies/reaction)with linearity over a 5-log_(10)-unit range in total hiv DNA assay.For the integrated hiv DNA assay,the LLOD was 8.0 copies/reaction(95%CI:5.8-16.6 copies/reaction)with linearity over a 3-log 10-unit range.Total hiv DNA in CD4^(+)T cells was positively associated with integrated hiv DNA(r=0.76,Phiv DNA and integrated hiv DNA in CD4^(+)T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8^(+)T-cell counts.Conclusions:This ddPCR assay can quantify total hiv DNA and integrated hiv DNA efficiently with robustness and sensitivity.It can be readily adapted for measuring hiv DNA with non-B clades,and it could be beneficial for testing in clinical trials.

关键词： Human immunodeficiency virus hiv Integrated hiv-1 DNA Total hiv DNA Droplet digital polymerase chain reaction hiv reservoir Antiretroviral therapy

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Temporal and Spatial Trends in hiv Positivity Rate for VCT Clinics—China,2015–2022

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China CDC weekly 2024年第1期6卷 6-11页

作者： Yi Liu Rong Su Dongmin Li Shaorong Wang Mengjie Han National center for AIDS/STD Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina

Introduction:Human immunodeficiency virus(hiv)voluntary counseling and testing(VCT)clinics play a critical role in identifying and diagnosing hiv cases.This study aimed to describe the trend of hiv positivity rate(HPR)among Chinese VCT clinics between 2015 and 2022.Methods:This study utilized data from the China Information System for Disease Control and Prevention to analyze the trend in the HPR for VCT clinics from 2015 to 2022.The HPR was calculated by dividing the number of newly-reported hiv cases by the number of hiv tests,multiplied by 100%.To identify temporal and spatial trends in the HPR,we employed joinpoint regression analysis and the Getis-Ord hotspot analysis.Results:From 2015 to 2022,VCT clinics in China performed a total of 22,075,386 hiv tests,leading to the identification of 260,353 hiv cases,resulting in a HPR of 1.18%.The HPR consistently declined over the study period,with an average annual percent change(AAPC)of−7.5%(95%confidence interval:−12.6%,−2.2%,Phiv cases.The declining HPR observed in these clinics may indicates the progress has been made in some degree in mitigating hiv among high-risk populations.Therefore,it is crucial to further improving the utilization of VCT clinics for hiv testing.

关键词： VCT dividing Spatial

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hiv Infection and Associated Factors Among Out-of-School MSM Aged 16–24 Years—6 Cities and Tianjin Municipality,China,2022–2023

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China CDC weekly 2024年第3期6卷 45-50页

作者： Hui Liu Qiyu Zhu Lei Zhang Yujing Liu Peng Xu Mengjie Han National center for AIDS/STD Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina Chinese Association of STD and AIDS Prevention and Control BeijingChina Beijing Chaoyang Kangzhong Health and Education center BeijingChina

What is already known about this topic?Approximately 80%of newly diagnosed human immunodeficiency virus(hiv)cases among individuals aged 15–24 years in China are attributed to out-ofschool youth.What is added by this report?This study presents findings on hiv prevalence and comprehensive insights into hiv service utilization,risky behaviors,and prevention knowledge among young out-of-school men who have sex with men(MSM)aged 16–24 years in China.This population exhibits a disproportionately high burden of hiv,with only 51.6%of hiv cases previously diagnosed.What are the implications for public health practice?hiv services should be expanded to include these key populations.Tailored interventions are needed to remove barriers to regular hiv services faced by young key populations.

关键词： prevention remove expanded

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Advances in technology for the laboratory diagnosis of individuals with hiv/AIDS coinfected with Mycobacterium tuberculosis

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生物安全与健康(英文) 2024年第3期6卷 133-142页

作者： Sun Jin Han Xiaoxu Yan Hongxia Zhang Xin Jiang Taiyi Zhang Tong Wu Hao Kaminskiy Grigory Ma Yingmin Karamov Eduard Su Bin Beijing Key Laboratory for hiv/AIDS Research Clinical and Research Center for Infectious Diseases Beijing Youan Hospital Capital Medical University Beijing China State Health Organization Tula Regional center for Control and Prevention of AIDS and Infectious Diseases Tula Russia Department of Respiratory and Critical Care Medicine Beijing Youan Hospital Capital Medical University Beijing China Gamaleya National Research Centre for Epidemiology and Microbiology of the Ministry of Health of the Russian Federation Moscow Russia

The high morbidity and mortality rate of individuals with human immunodeficiency virus (hiv) / acquired immunodeficiency syndrome (AIDS) coinfected withMycobacterium tuberculosis (MTB) is a tough challenge for current global tuberculosis prevention and control efforts. hiv/MTB coinfection is more complex than a single infection, and the interaction between the two diseases aggravates the deterioration caused by the disease, resulting in increased hospitalizations and deaths. Rapid screening and early diagnosis facilitate the timely initiation of anti-tuberculosis treatment in hiv/MTB coinfected individuals, thereby reducing transmission and the incidence of adverse prognoses. To date, pathogenic detection has remained the gold standard for diagnosing tuberculosis, but its sensitivity and specificity are greatly affected by the body''s immune status, which limits its application in the diagnosis of hiv/MTB coinfection. Recently, immunology and molecular detection technology has developed rapidly. New detection technologies, such as interferon-γ release assays, interferon-gamma inducible protein 10, and GeneXpert MTB/RIF assay have overcome the limitations of traditional detection methods, significantly improved the sensitivity and specificity of tuberculosis diagnosis, and brought new hope to the detection of hiv/MTB coinfection. In this article, the principle, scope of application, and latest research progress of relevant detection methods are reviewed to provide a reference for the early diagnosis of hiv/MTB coinfection.

关键词： Human immunodeficiency virus (hiv)/Mycobacterium tuberculosis (MTB) coinfection Mycobacterium tuberculosis (MTB) Early diagnosis

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Characteristics of the hiv/AIDS Epidemic among People Aged≥50 Years in China during 2018–2021

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Biomedical and Environmental Sciences 2024年第4期37卷 399-405页

作者： HOU Yu Shan JIN Yi Chen CAI Chang TANG Hou Lin QIN Qian Qian LYU Fan National center for AIDS/STD Control and Prevention Chinese Center for Disease Control and PreventionBeijing 102206China

Objective This study aimed to determine the current epidemiological status of PLWHA aged≥50 years in China from 2018 to 2021.It also aimed to recommend targeted interventions for the prevention and treatment of hiv/AIDS in elderly patients.Methods Data on newly reported cases of PLWHA,aged≥50 years in China from 2018 to 2021,were collected using the CRIMS.Trend tests and spatial analyses were also conducted.Results Between 2018 and 2021,237,724 hiv/AIDS cases were reported among patients aged≥50 years in China.The main transmission route was heterosexual transmission(91.24%).Commercial heterosexual transmission(CHC)was the primary mode of transmission among males,while non-marital non-CHC([NMNCHC];60.59%)was the prevalent route in women.The proportion of patients with CHC decreased over time(Z=67.716,Phiv/AIDS cases exhibited spatial clustering.Based on these results,the expansion of hiv testing,treatment,and integrated behavioral interventions in high-risk populations is recommended to enhance disease detection in key regions.

关键词： hiv/AIDS Aged≥50 years China

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Overexpression of wild-type HRAS drives non-alcoholic steatohepatitis to hepatocellular carcinoma in mice

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Zoological Research 2024年第3期45卷 551-566页

作者： Chen Ling Su-Su Liu Yu-Ya Wang Gui-Tao Huo Yan-Wei Yang Nan Xu Hong Wang Yong Wu Yu-Fa Miao Rui Fu Yu-Wei Zhao Chang-Fa Fan College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceXi’anShaanxi 710069China Division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China National center for Safety Evaluation of Drugs National Institutes for Food and Drug Control(NIFDC)Beijing 100176China Division of hiv/AIDS and Sexually Transmitted Virus Vaccines Institute for Biological Products ControlNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China Division of Laboratory Animal Monitoring Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China

Hepatocellular carcinoma(HCC),a prevalent solid carcinoma of significant concern,is an aggressive and often fatal disease with increasing global incidence rates and poor therapeutic outcomes.The etiology and pathological progression of non-alcoholic steatohepatitis(NASH)-related HCC is multifactorial and multistage.However,no single animal model can accurately mimic the full NASH-related HCC pathological progression,posing considerable challenges to transition and mechanistic studies.Herein,a novel conditional inducible wild-type human HRAS overexpressed mouse model(HRAS-HCC)was established,demonstrating 100%morbidity and mortality within approximately one month under normal dietary and lifestyle conditions.Advanced symptoms of HCC such as ascites,thrombus,internal hemorrhage,jaundice,and lung metastasis were successfully replicated in mice.In-depth pathological features of NASH-related HCC were demonstrated by pathological staining,biochemical analyses,and typical marker gene detections.Combined murine anti-PD-1 and sorafenib treatment effectively prolonged mouse survival,further confirming the accuracy and reliability of the model.Based on protein-protein interaction(PPI)network and RNA sequencing analyses,we speculated that overexpression of HRAS may initiate the THBS1-COL4A3 axis to induce NASH with severe fibrosis,with subsequent progression to HCC.Collectively,our study successfully duplicated natural sequential progression in a single murine model over a very short period,providing an accurate and reliable preclinical tool for therapeutic evaluations targeting the NASH to HCC continuum.

关键词： HRAS THBS1 HCC driver factor NASH Fibrosis Cirrhosis HCC Treatment evaluation

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Current status of liver transplantation for human immunodeficiency virus-infected patients in China's Mainland

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World Journal of Gastroenterology 2024年第14期30卷 1958-1962页

作者： Jian-Xin Tang Dong Zhao Department of Liver Surgery and Organ Transplantation center Shenzhen Third People’s HospitalThe Second Affiliated Hospital of Southern University of Science and TechnologyNational Clinical Research Center for Infectious DiseaseShenzhen 518000Guangdong ProvinceChina

According to the report from the Chinese center for Disease Control and Prevention,the prevalence of human immunodeficiency virus(hiv)infection exceeded 1.2 million individuals by the year 2022,with an annual increase of about 80000 cases.The overall prevalence of hepatitis B surface antigen among individuals co-infected with hiv reached 13.7%,almost twice the rate of the general population in China.In addition to the well-documented susceptibility to opportunistic infections and new malignancies,hiv infected patients frequently experience liver-related organ damage,with the liver and kidneys being the most commonly affected.This often leads to the development of end-stage liver and kidney diseases.Therefore,organ transplantation has emerged as an important part of active treatment for hiv infected patients.However,the curative effect is not satisfactory.hiv infection has been considered a contraindication for organ transplantation.Until the emergence of highly active anti-retroviral therapy in 1996,the once intractable replication of retrovirus was effectively inhibited.With prolonged survival,the failure of important organs has become the main cause of death among hiv patients.Therefore,transplant centers worldwide have resu-med exploration of organ transplantation for hiv-infected individuals and reached a positive conclusion.This study provides an overview of the current landscape of hiv-positive patients receiving liver transplantation(LT)in main-land China.To date,our transplant center has conducted LT for eight end-stage liver disease patients co-infected with hiv,and all but one,who died two months postoperatively due to sepsis and progressive multi-organ failure,have survived.Comparative analysis with hepatitis B virus-infected patients during the same period revealed no statistically significant differences in acute rejection reactions,cytomegalovirus infection,bacteremia,pulmonary infections,acute kidney injury,new-onset cancers,or vascular and biliary complications.

关键词： Liver transplantation Human immunodeficiency virus Infection Hepatitis B virus End-stage liver disease China'Mainland

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DNA framework signal amplification platform-based highthroughput systemic immune monitoring

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Signal Transduction and Targeted Therapy 2024年第3期9卷 1115-1131页

作者： Ye Chen Xingyu Chen Bowen Zhang Yuxin Zhang Songhang Li Zhiqiang Liu Yang Gao Yuxuan Zhao Lin Yan Yi Li Taoran Tian Yunfeng Lin State Key Laboratory of Oral Diseases National Center for StomatologyNational Clinical Research Center for Oral DiseasesWest China Hospital of StomatologySichuan UniversityChengdu 610041 SichuanPR China Department of Prosthodontics Tianjin Stomatological HospitalSchool of MedicineNankai UniversityTianjin 300041PR China Department of Laboratory Medicine West China HospitalSichuan UniversityChengdu 610041 SichuanPR China Sichuan Provincial Engineering Research center of Oral Biomaterials Chengdu 610041 SichuanChina

Systemic immune monitoring is a crucial clinical tool for disease early diagnosis,prognosis and treatment planning by quantitative analysis of immune cells.However,conventional immune monitoring using flow cytometry faces huge challenges in large-scale sample testing,especially in mass health screenings,because of time-consuming,technical-sensitive and high-cost features.However,the lack of high-performance detection platforms hinders the development of high-throughput immune monitoring technology.To address this bottleneck,we constructed a generally applicable DNA framework signal amplification platform(DSAP)based on post-systematic evolution of ligands by exponential enrichment and DNA tetrahedral framework-structured probe design to achieve high-sensitive detection for diverse immune cells,including CD4+,CD8+T-lymphocytes,and monocytes(down to 1/100μl).Based on this advanced detection platform,we present a novel high-throughput immune-cell phenotyping system,DSAP,achieving 30-min one-step immune-cell phenotyping without cell washing and subset analysis and showing comparable accuracy with flow cytometry while significantly reducing detection time and cost.As a proof-of-concept,DSAP demonstrates excellent diagnostic accuracy in immunodeficiency staging for 107 hiv patients(AUC>0.97)within 30 min,which can be applied in hiv infection monitoring and screening.Therefore,we initially introduced promising DSAP to achieve high-throughput immune monitoring and open robust routes for point-of-care device development.

关键词： diagnosis monitoring initially

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Knowledge,Attitude,and Practice Towards Mpox and Associated Factors Among hiv-Infected Individuals—Beijing Municipality,China,2023

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China CDC weekly 2024年第7期6卷 109-117,I0001,I0002页

作者： Yue Gu Ruiqi Ren Jing Han Wenqing Bai Yuanyuan Zhang Haoliang Liu Zhaohe Li Chao Li Rui Song Lei Zhou Chinese center for Disease Control and Prevention BeijingChina National center for Infectious Diseases Beijing Ditan HospitalCapital Medical UniversityBeijingChina Beijing Key Laboratory of Emerging Infectious Diseases Institute of Infectious DiseasesBeijing Ditan HospitalCapital Medical UniversityBeijingChina Nanjing Medical University Nanjing CityJiangsu ProvinceChina

Summary What is already known about this topic?Approximately 50%of patients with mpox are human immunodeficiency virus(hiv)-infected globally.Studies have shown that individuals with advanced hiv infection tend to hav... 详细信息

关键词： mortality Infect clinical

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A Case of Acute hiv-1 and Monkeypox Coinfection After Condomless Insertive Anal Sex in the Previous 69 Days—Beijing Municipality,China,August-October,2023

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China CDC weekly 2024年第7期6卷 126-130页

作者： Lin Jia Han Jia Yuan Fang Benyong Yan Mei Zhang Yang Zhang Wen Wang Caiping Guo Xiaojie Huang Tong Zhang Taiyi Jiang Clinical and Research center for Infectious Diseases Beijing Youan HospitalCapital Medical UniversityBeijingChina

Summary What is already known about this topic?The prevalence of monkeypox(mpox)infections is primarily observed among young men who engage in sexual activities with other men,and there is a possibility of sexual transmission.Co-occurring sexually transmitted infections have also been documented.What is added by this report?In this report,we present a case of a patient in China who was simultaneously diagnosed with mpox,and acute human immunodeficiency virus(hiv)infection.

关键词： acute Acute prevalence

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