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A New High-throughput Real-time PCR Assay for the Screening of Multiple Antimicrobial Resistance Genes in Broiler Fecal Samples from China

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Biomedical and Environmental Sciences 2019年第12期32卷 881-892页

作者： CHE Jie LU Jin Xing LI Wen Ge ZHANG Yun Fei ZHAO Xiao Fei YUAN Min BAI Xue Mei CHEN Xia LI Juan state key laboratory of infectious Disease prevention and control Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention

Objective Antimicrobial resistance(AMR)has become a global concern and is especially severe in *** effectively and reliably provide AMR data,we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology,and screened multiple AMR genes in broiler fecal *** A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection.A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR *** The new assay with limits of detection ranging from 40.9 to 8,000 copies/*** sensitivity rate,specificity rate,positive predictive value,negative predictive value and correct indices were 99.30%,98.08%,95.31%,99.79%,and 0.9755,*** this assay,we demonstrate that AMR genes are widely spread,with positive detection rates ranging from 0 to 97.07%in 273 broiler fecal *** CTX-M,bla TEM,mcr-1,fex A,cfr,optr A,and int I1 showed over 80%*** dissemination of AMR genes was distinct between the two *** We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal *** widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.

关键词： Antimicrobial resistance Antimicrobial resistance gene High-throughput real-time PCR array Broiler fecal sample

Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress

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Biomedical and Environmental Sciences 2018年第12期31卷 867-875页

作者： LUO Xiao Nuan YAO Hai Lan SONG Juan SONG Qin Qin SHI Bing Tian XIA Dong HAN Jun state key laboratory of infectious Disease prevention and control Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Molecular Immunology laboratory Capital Institute of Pediatrics

Objective Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3(CVB3) infection induced autophagy through endoplasmic reticulum(ER) stress. Methods In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction(RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase(PERK) inhibitor, inositol-requiring protein-1(IRE1) inhibitor, or activating transcription factor-6(ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3(LC3). Results CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein(GFP)-LC3 punctuation and induced the conversion from LC3-Ⅰ to phosphatidylethanolamine-conjugated LC3-1(LC3-Ⅱ). CVB3 infection still decreased the expression of mammalian target of rapamycin(mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-Ⅱ to LC3-Ⅰ in CVB3-infected HeLa cells. Conclusion CVB3 infection induced autophagy through ER stress in HeL a cells, and PERK, IRE1, and ATF6 a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.

关键词： Coxsackievirus B3(CVB3) Autophagy Endoplasmic reticulum(ER) stress Unfolded protein response(UPR)

Preliminary Study on Drug Susceptibility Profile and Resistance Mechanisms to Macrolides of Clinical Isolates of Non-tuberculous Mycobacteria from China

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Biomedical and Environmental Sciences 2018年第4期31卷 290-299页

作者： LI Fu LI Gui Lian PANG Hui LIU Hai Can XIAO Tong Yang LI Shuang Jun LUO Qiao JIANG Yi WANG Rui Bai WAN Kang Lin state key laboratory for infectious Diseases prevention and control Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention Immunology Department Changzhi Medical College key laboratory of laboratory Medicine Wenzhou Medical University

Objective Macrolide susceptibility and drug resistance mechanisms of clinical non-tuberculous mycobacteria（NTM） isolates were preliminarily investigated for more accurate diagnosis and treatment of the infection in China. Methods Four macrolides, including clarithromycin（CLAR）, azithromycin（AZM）, roxithromycin（ROX）, and erythromycin（ERY）, were used to test the drug susceptibility of 310 clinical NTM isolates from six provinces of China with the broth microdilution method. Two resistance mechanisms, 23 S r RNA and erm, were analyzed with nucleotide sequence analysis. Results Varied effectiveness of macrolides and species-specific resistance patterns were observed. Most Mycobacterium abscessus subsp. massiliense were susceptible and all M. fortuitum were highly resistant to macrolides. All the drugs, except for erythromycin, exhibited excellent activities against slow-growing mycobacteria, and drug resistance rates were below 22.2%. Only four highly resistant strains harbored 2,058/2,059 substitutions on rrl and none of other mutations were related to macrolide resistance. G2191 A and T2221 C on rrl were specific for the M. abscessus complex（MABC）. Seven sites, G2140 A, G2210 C, C2217 G, T2238 C, T2322 C, T2404 C, and A2406 G, were specifically carried by M. avium and M. intracellulare. Three sites, A2192 G, T2358 G, and A2636 G, were observed only in M. fortuitum and one site G2152 A was specific for M. gordonae. The genes erm（39） and erm（41） were detected in M. fortuitum and M. abscessus and inducible resistance was observed in relevant sequevar. Conclusion The susceptibility profile of macrolides against NTM was demonstrated. The well-known macrolide resistance mechanisms, 23 S r RNA and erm, failed to account for all resistant NTM isolates, and further studies are warranted to investigate macrolide resistance mechanisms in various NTM species.

关键词： Non-tuberculous mycobacteria Macrolide Drug resistance

Impact of HIV Pretreatment Drug Resistance on Virological Failure After One-Year Antiretroviral Therapy--China,2018-2019

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China CDC weekly 2022年第25期4卷 535-540页

作者： Miaomiao Li Chang Song Aobo Dong Jing Hu Ruihua Kang Hui Xing Yuhua Ruan Yiming Shao Kunxue Hong Lingjie Liao state key laboratory of infectious Disease prevention and control National Center for AIDS/STD Control and PreventionChinese Center for Disease Control and PreventionBeijingChina

Summary What is already known about this topic?While antiretroviral therapy(ART)has been rapidly scaled-up among the population living with human immunodeficiency virus or acquired immune deficiency syndrome(HIV/AIDS)... 详细信息

关键词： Therapy Impact patients

Trans-Regional and Cross-Host Spread of mcr-Carrying Plasmids Revealed by Complete Plasmid Sequences——44 Countries,1998−2020

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China CDC weekly 2022年第12期4卷 242-248,I0001-I0004页

作者： Zhe Li Zhenpeng Li Yao Peng Xin Lu Biao Kan state key laboratory of infectious Disease prevention and control/National Institute for Communicable Disease control and prevention Chinese Center for Disease Control and PreventionBeijingChina

Background:The surveillance of antimicrobial resistance genes(ARGs)and bacteria is one critical approach to prevent and control antimicrobial resistance(AMR).Next-generation sequencing(NGS)is a powerful tool in monitoring the emergence and spread of ARGs and resistant *** horizontal transfer of ARGs across host bacteria mediated by plasmids is a challenge in NGS surveillance for resistance because short-read sequencing can hardly generate the complete plasmid genome sequence,and the correlation between ARGs and plasmids are difficult to ***:The complete genome sequences of 455 mcr-carrying plasmids(pMCRs),and the data of their host bacteria and isolation regions were collected from the NCBI *** of Inc types and ARGs were searched for each *** genome similarity of these plasmids was analyzed by pangenome clustering and genome ***:A total of 52 Inc types,including a variety of fusion plasmids containing 2 or more Inc types were identified in these pMCRs and carried by complex host *** cooccurrence of ARGs in pMCRs was generally observed,with an average of 3.9 ARGs per ***-two clusters with consistent or highly similar sequences and gene compositions were identified by the pangenome clustering,which were characterized with distributions in different countries/regions,years or host bacteria in each ***:Based on the complete plasmid sequences,distribution of mcr genes in different Inc type plasmids,their co-existence with other AMRs,and transmission of one pMCR across regions and host bacteria can be revealed *** plasmid genomes and comparisons in the laboratory network are necessary for spread tracing of ARG-carrying plasmids and risk assessment in AMR surveillance.

关键词： sequences resistance Host

Development and application of a real-time polymerase chain reaction method for Campylobacter jejuni detection

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World Journal of Gastroenterology 2013年第20期19卷 3090-3095页

作者： Mao-Jun Zhang Bo Qiao Xue-Bin Xu Jian-Zhong Zhang state key laboratory for infectious Disease prevention and control and National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and Prevention Shanghai Municipal Center for Disease control and prevention

AIM:To develop a real-time polymerase chain reaction(PCR) method to detect and quantify Campylobacter jejuni(***) from stool ***:Primers and a probe for real-time PCR were designed based on the specific DNA sequence of the hipO gene in *** specificity of the primers and probe were tested against a set of Campylobacter *** other enteric *** optimal PCR conditions were determined by testing a series of conditions with standard a *** *** detection limits were obtained using purified DNA from bacterial culture and extracted DNA from the stool *** hundred and forty-two specimens were analyzed for the presence of *** by direct bacterial culture and real-time ***:The optimal PCR system was determined using reference DNA templates,1 × uracil-DNA glycosylase,3.5 mmol/L MgCl 2,1.25 U platinum Taq polymerase,0.4 mmol/L PCR nucleotide mix,0.48 μmol/L of each primer,0.2 μmol/L of probe and 2 μL of DNA template in a final volume of 25 μ*** PCR reaction was carried as follows:95 ℃ for 4 min,followed by 45 cycles of 10 s at 95 ℃ and 30 s at 59 ℃.The detection limit was 4.3 CFU/mL using purified DNA from bacterial culture and 10 3 CFU/g using DNA from stool ***(8.3%,20/242) *** strains were isolated from bacterial culture,while 41(16.9%,41/242) samples were found to be positive by realtime *** sequencing of the PCR product indicated the presence of *** in the *** mixed infection of *** and Salmonella was detected in one specimen and the PCR test for this specimen was ***:The sensitivity of detection of *** from stool specimens was much higher using this PCR assay than using the direct culture method.

关键词： Campylobacter jejuni Real time polymerase chain reaction Application

Role of the blood-brain barrier in rabies virus infection and protection

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Protein & Cell 2013年第12期4卷 901-903页

作者： Lihua Wang Yuxi Cao Qing Tang Guodong Liang state key laboratory for infectious Disease prevention and control Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China

Rabies is an acute,progressive encephalitis caused by infection with rabies virus(RABV).It is one of the most important zoonotic infections and causes more than 70,000 human deaths annually(http://***).It has long been held that a rabies infection is lethal in humans once the causative RABV reaches the central nervous system(CNS);however,this concept was challenged by the recent recovery of a small number of rabies *** analysis of these patients revealed that the blood-brain barrier(BBB)played a major role in protection against the *** main reason for the survival of these patients was enhanced BBB permeability after infection with the causative agent(usually bat-originated RABV showing reduced pathogenicity),which allowed immune cells to enter the tissues of the CNS and clear the infection(Willoughby et al.,2005).These fi ndings have been confi rmed in animal infection experiments(Wang et al.,2005;Roy and Hooper,2007,2008;Faber et al.,2009).Thus,the BBB has attracted the attention of scientists interested in the pathogenesis of,and therapeutic approaches,for *** paper introduces the role of the BBB in rabies infections and protection of the CNS and provides insight into future treatments for patients with clinical rabies.

关键词： rabies infection barrier

Changes in HIV-1 Subtypes/Sub-Subtypes,and Transmitted Drug Resistance Among ART-Naïve HIV-Infected Individuals—China,2004–2022

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China CDC weekly 2023年第30期5卷 664-671,I0010-I0012页

作者： Xiu Liu Dong Wang Jing Hu Chang Song Lingjie Liao Yi Feng Dan Li Hui Xing Yuhua Ruan state key laboratory of infectious Disease prevention and control(SKLID) National Center for AIDS/STD Control and PreventionChinese Center for Disease Control and Prevention(China CDC)Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesBeijingChina.

Introduction:The efficacy of treatment and clinical outcomes may be jeopardized by factors such as transmitted drug resistance(TDR)and the genetic diversity of the human immunodeficiency virus type 1(HIV-1).This comprehensive study aims to examine the alterations in HIV-1 subtypes or sub-subtypes and TDR among Chinese individuals,who have been diagnosed with HIV infection and are previously untreated with antiretroviral therapy(ART),across the span of 2004 to ***:Sequences of the HIV-1 pol gene region were obtained from ART-naïve HIV-positive individuals across 31 provincial-level administrative divisions between 2004 and *** predict susceptibility to 12 antiretroviral drugs,the research utilized the Stanford HIV Drug Resistance *** Cochran-Armitage trend test facilitated the analysis of changes in HIV-1 subtype/sub-subtype prevalence and *** analysis was conducted in alignment with the progression of the National Free Antiretroviral Treatment Program’s stages between 2004 and ***:Among the 57,902 ART-naïve individuals infected with HIV,there was a notable decline in the prevalence of CRF01_AE,B,and C from 37.3%,24.1%,and 1.3%respectively in 2004–2007 to 29.4%,7.3%,and 0.2%respectively in 2020–***,a significant increase was observed in the proportions of CRF07_BC,CRF08_BC,CRF55_01B,other CRFs,and URFs,from 24.1%,11.5%,0.1%,0.4%,and 0.9%respectively in 2004–2007 to 40.8%,11.5%,3.8%,3.7%,and 2.8%respectively in 2020–2022(all P<0.001 for trend).The prevalence of TDR to overall,non-nucleoside reverse transcriptase inhibitor(NNRTI),efavirenz,and nevirapine also significantly increased from 2.6%,1.8%,1.6%,and 1.8%respectively in 2004–2007 to 7.8%,6.7%,6.3%,and 6.7%respectively in 2020–2022(all P<0.001 for trend).However,there were no meaningful changes in the TDR prevalence of nucleoside reverse transcriptase inhibitor and protease ***,in 2020–2022,the overall TDR prevalence exceeded 15%in ***

关键词： prevalence alterations drugs

Comparison of the Pathogenicity of Neisseria meningitidis Isolates of Hyperinvasive Sequence Type 7 Belonging to Serogroups A,B,C,and X

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Biomedical and Environmental Sciences 2020年第2期33卷 114-122页

作者： GUO Peng Bo ZHU Bing Qing XU Li GAO Wan Ying GAO Yuan LIANG Hao ZHANG Mao Jun SHAO Zhu Jun state key laboratory for infectious Disease prevention and control National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Collaborative Innovation Center for Diagnosis and Treatment of infectious Diseases Hangzhou 310003ZhejiangChina

Objective To compare the pathogenicity of isolates of sequence type 7(ST-7)Neisseria meningitidis(***)belonging to four different serogroups(A,B,C,and X).Methods Four ST-7 *** isolates serogrouped as A,B,C,and X and characterized by different capsule structures,were examined for their adhesion and invasion properties,and their ability to induce cytokine release and apoptosis in the host cell(the A549 cell line).Results Among the four ST-7 *** isolates,the serogroup A isolate possessed the strongest adhesion and invasion *** isolate also induced the release of the highest levels of the proinflammatory mediators interleukin-6,interleukin-1β,and interferon,and the highest apoptosis rate in the host ***,there was no significant difference in interleukin-8 and tumor necrosis factor-αsecretion between the four *** on the findings,the serogroup X *** isolate had the weakest pathogenicity,whereas there was almost no difference in the pathogenicity of the isolates from serogroups B and *** The differences in the capsular structure of the four isolates of ST-7 *** affected their pathogenic *** findings also imply that the hyperinvasive ST-7 *** lineage may include hypoinvasive isolates.

关键词： ST-7 N.meningitidis Capsule Pathogenicity Adaptation