耐碳青霉烯类鲍曼不动杆菌OXA和NDM-1耐药基因的研究

目的检测江苏盛泽医院耐碳青霉烯类抗生素鲍曼不动杆菌的OXA和NDM-1耐药基因,分析耐碳青霉烯类抗菌药物的耐药机制。方法采用改良Hodge试验检测30株耐碳青霉烯类抗生素鲍曼不动杆菌产酶情况;用PCR的方法检测OXA-23、OXA-24、VIM、IMP和NDM-1碳青霉烯酶耐药基因。结果 30株分离菌中25株菌改良Hodge试验阳性,22株携带OXA-23型碳青霉烯酶耐药基因,未扩增出NDM-1碳青霉烯酶耐药基因。结论本院耐碳青霉烯类抗生素鲍曼不动杆菌的耐药机制主要是携带OXA-23型碳青霉烯酶基因。     

鲍曼不动杆菌的耐药情况及碳青霉烯酶基因检测

了解佳木斯大学附属第一医院鲍曼不动杆菌耐药性及碳青霉烯酶包括苯唑西林酶和金属酶相关耐药基因分布情况,为临床抗菌药物的合理选择提供依据。2013年9月至2014年12月使用VITEK-II全自动微生物鉴定/药敏测试系统筛选出佳木斯大学附属第一医院临床标本鲍曼不动杆菌69株;采用多重PCR方法检测鲍曼不动杆菌携带的碳青霉烯酶相关耐药基因16SrRNA、OXA-23、OXA-24、OXA-51、OXA-58、IMP、VIM、SIM,并对耐药基因扩增的阳性产物进行DNA 序列分析。69株AB对亚胺培南、美洛培南的耐药率分别为36.2%、37.68%,对其他抗菌药物的耐药率均高于50%。6种耐药基因的检测结果为69株(100%)携带OXA-51基因,32株(46.4%)携带OXA-23基因,17株(24.6%)携带OXA-24基因,5株(7.2%)携带OXA 58基因,1株（1.4%）携带IMP基因。25株碳青霉烯类药物耐药鲍曼不动杆菌中,22株(88%) 携带OXA-23,1株(4%)携带OXA-58,10株(40%)携带OXA-24,6株（24%）同时携带OXA-23、OXA-24。 DNA序列分析结果显示：OXA-23、OXA-24、OXA-51、OXA-58分别与NCBI的序列同源性均为99%。产OXA-23型碳青霉烯酶可能是佳木斯大学附属第一医院鲍曼不动杆菌对碳青霉烯酶类抗菌药物耐药的主要原因,另外佳木斯大学附属第一医院存在OXA-24型耐药基因鲍曼不动杆菌的区域性流行。     

鲍曼不动杆菌对碳青霉烯类抗生素的耐药性及其基因的分析

目的研究23株鲍曼不动杆菌对碳青霉烯类抗生素的耐药情况及对耐药基因分析,为临床用药提供依据。方法用珠海迪尔DL-96鉴定系统进行细菌鉴定及K-B法进行药敏试验,用碳青霉烯酶4种基因的特异性引物对其进行聚合酶链反应(PCR)扩增和基因型分析,并通过网上GenBank进行比对以确定编码酶基因的类型。结果 23株鲍曼不动杆菌对哌拉西林/他唑巴坦、左旋氧氟沙星、丁胺卡那霉素、多黏菌素B的耐药率分别为80%、45%、30%、10%,对其他抗生素的耐药率均在90%以上。携带D类碳青霉烯酶OXA-23基因有18株(78%),携带OXA-51基因有15株(65%),OXA-24、OXA-58基因引物PCR扩增为阴性,随机各抽取3株OXA-23基因阳性株进行测序后通过在网上GenBank比对与OXA-23标准株99%同源,OXA-51基因阳性株与OXA-51标准株98%同源。结论耐碳青霉烯类抗生素的鲍曼不动杆菌对多黏菌素的耐药率最低,其次是丁胺卡那霉素,其中以携带OXA-23型碳青霉烯酶基因为主,应引起临床高度关注,防止在院内广泛传播。     

耐碳青霉烯类鲍曼不动杆菌OXA基因检测及同源性分析

目的 了解台州地区碳青霉烯类耐药鲍曼不动杆菌的耐药性、碳青霉烯酶基因型以及同源性.方法 63株碳青霉烯类耐药鲍曼不动杆菌经VITEK2 Compact进行细菌鉴定及药敏分析,用K-B法复核药敏结果,采用多重PCR扩增分析碳青霉烯酶的基因型;采用脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)分析其同源性.结果 63株菌株为多重耐药菌株,除多粘菌素、阿米卡星、头孢哌酮/舒巴坦外,对其他常用抗生素的耐药率均在70％以上.63株菌株检测出OXA-51基因60株(95.2％),OXA-23基因58株(92.1％),两个基因同时存在的有58株(92.1％).PFGE结果显示碳青霉烯类鲍曼不动杆菌主要分为5个克隆型,其中A、B两个为主型.结论 产OXA酶是台州地区耐碳青霉烯类鲍曼不动杆菌的主要耐药机制之一,其中OXA-23是主要的基因型.     

耐碳青霉烯鲍曼不动杆菌 插入序列及其水平传播

目的了解我院耐碳青霉烯类鲍曼不动杆菌的临床分布并探讨插入序列与其耐药的关系,分析水平传播能力,为指导医院感染及临床合理应用抗菌药物提供科学依据。方法收集2013年9月-2015年6月我院临床分离鲍曼不动杆菌,经VITEK-II全自动细菌分析系统鉴定细菌并检测16SrRNA,Walkway-40/药敏测试系统进行药敏检测;多重PCR检测鲍曼不动杆菌携带β-内酰胺酶(A、B、C、D类)相关耐药基因。检测上游插入序列ISAba1与OXA-23、OXA-51、ADC连锁表达,并分析ISAbal与耐药基因OXA-23、ADC的相关性。质粒接合试验验证OXA碳青霉烯酶基因的水平转移。结果耐碳青霉烯类的鲍曼不动杆菌(CRAB)与碳青霉烯类敏感的鲍曼不动杆菌(CSAB)抗生素耐药率差异有统计学意义(Ps0.01)。CRAB与CSAB产酶基因(OXA-23、ADC、TEM)检出率差异明显。50株CRAB中40株检测出ISAbal-OXA-23连锁基因,1株检测出ISAbal-OXA-51连锁基因。接合试验阳性株检测出OXA-23、OXA-24、OXA-51及插入序列。结论我院CRAB主要是产OXA-23、OXA-24、OXA-51、ADC、TEM型碳青霉烯酶,ISAbal常出现在OXA-23基因上游,ISAbal-OXA-23可能是CRAB重要的耐药机制。     

鲍曼不动杆菌中OXA碳青霉烯酶的表型筛选与PCR检测

摘要：目的 了解OXA碳青霉烯酶在暨南大学附属第一医院耐亚胺培南鲍曼不动杆菌中的流行状况,完善OXA碳青霉烯酶的分子流行病学资料。方法 采用改良Hodge试验筛选产碳青霉烯酶菌株,多重PCR法检测OXA碳青霉烯酶的编码基因（blaOXA-23-like、blaOXA-24-like、blaOXA-58-like和blaOXA-143）,利用生物信息学的方法对OXA亚型进行比对分析并制作分子进化树。结果 在157株耐亚胺培南鲍曼不动杆菌中Hodge试验筛选出碳青霉烯酶表型阳性菌株141株,PCR检测结果显示有132株携带OXA-23编码基因,未检测到OXA-24-like、OXA-58-like和OXA-143亚型。结论 产OXA-23碳青霉烯酶是该院鲍曼不动杆菌对亚胺培南耐药的主要机制之一。     

危重病房耐碳青霉烯酶鲍曼不动杆菌同源性研究

目的探讨杭州市第一医院危重病房耐碳青霉烯酶鲍曼不动杆菌之间的同源性,进行分子流行病学调查,旨在为制定预防和控制其院内感染的措施提供依据。方法收集该院危重病房2005年1月至12月分离到的34株亚胺培南耐药鲍曼不动杆菌。采用全自动微生物分析系统VITEK-AMS60对34株耐碳青霉烯酶鲍曼不动杆菌进行鉴定及药敏;用琼脂稀释法和E-test法测定14种抗菌药物的最低抑菌浓度(MIC),脉冲场凝胶电泳(PF-GE)分析其耐药株的同源性,对碳青霉烯类基因OXA-23型、OXA-24型、IMP型、VIM型基因进行PCR扩增及序列分析。结果PFGE发现34株鲍曼不动杆菌菌株为同一耐药克隆株,在危重病房呈爆发流行。所有对亚胺培南耐药鲍曼不动杆菌明确产OXA-23型碳青霉烯酶,未检出OXA-24、IMP、VIM基因型。34株菌株质粒提取未成功。结论该院同一个耐药克隆株在危重病房不同患者身上流行,可能与行气管插管、呼吸机、氧气湿化瓶、护士手操作有关。     

鲍曼不动杆菌对碳青霉烯类抗生素的耐药性分析

目的了解鲍曼不动杆菌的耐药情况,并检测耐碳青霉烯类鲍曼不动杆菌的耐药基因,为指导临床合理用药、控制院内感染提供依据。方法利用K-B法检测45株鲍曼不动杆菌临床分离株的耐药情况,通过改良Hodge试验、Carba NP试验和EDTA协同试验对多重耐药鲍曼不动杆菌的碳青霉烯酶进行表型检测,并采用PCR技术检测鲍曼不动杆菌携带OXA-23和NDM-1型耐药基因的情况。结果 45株鲍曼不动杆菌临床分离株中共筛出42株多重耐药菌株;利用改良Hodge试验和Carba NP试验检出36株碳青霉烯酶阳性菌株;采用PCR扩增出OXA-23,未扩增出NDM-1。结论鲍曼不动杆菌耐药情况严重,且耐药基因OXA-23携带率高,治疗时应根据药敏试验结果合理用药。     

29株耐亚胺培南和/或美罗培南鲍曼不动杆菌产碳青霉烯酶基因型检测

目的 了解临床分离耐亚胺培南和/或耐美罗培南鲍曼不动杆菌中产碳青霉烯酶的基因型别.方法 采用聚合酶链反应扩增IMP、VIM、OXA型碳青霉烯酶基因并测序.结果 29株对碳青霉烯类耐药的鲍曼不动杆菌中,以产OXA-24型和IMP型酶菌株最多,二者均占51.7％ (15/29).产OXA-24+ IMP型5株、OXA-24+ OXA-51+IMP型4株、VIM型4株、OXA-24+ OXA-58+ IMP型、OXA-23+ OXA-24+ IMP型各2株,OXA-23+IMP型、OXA-51+OXA-24型、OXA-24型、IMP型各1株,8株细菌PCR检测结果为阴性.结论 耐亚胺培南和/或美罗培南鲍曼不动杆菌主要产OXA-24型和IMP型碳青霉烯酶,部分菌株可同时产2种或以上碳青霉烯酶.     

黑龙江省东部地区鲍曼不动杆菌碳青霉烯酶基因检测

目的探讨鲍曼不动杆菌(Acinetobacter baumannii,A.baumannii)耐药性及碳青霉烯酶相关耐药基因OXA-23、OXA-24、OXA-51和OXA-58的分布情况,为临床抗菌药物的合理选择提供依据。方法 2014年1至月2014年12月收集佳木斯大学附属第一医院临床标本(包括痰、分泌物、脑脊液、血液、咽拭子等标本),使用VITEK-II全自动微生物鉴定/药敏测试系统筛选出44株A.baumannii;采用多重PCR检测A.baumannii携带的碳青霉烯酶相关耐药基因OXA-23、OXA-24、OXA-51、OXA-58,并对耐药基因扩增的阳性产物进行DNA序列分析。结果 44株A.baumannii对复方新诺明、亚胺培南、左氧氟沙星的敏感率分别为65.91%、61.36%、61.36%,对其他抗菌药物的敏感率均低于50.00%。4种耐药基因的检测结果为:44株(100.00%)携带OXA-51基因,20株(45.45%)携带OXA-23基因,14株(31.82%)携带OXA-24基因,3株(6.82%)携带OXA-58基因。16株碳青霉烯类药物耐药A.baumannii中,14株(87.50%)携带OXA-23,1株(6.25%)携带OXA-58,8株(50.00%)携带OXA-24,5株(31.25%)同时携带OXA-23、OXA-24。DNA序列分析结果显示:OXA-23、OXA-24、OXA-51、OXA-58分别与NCBI的序列同源性均为99.00%。结论A.baumannii耐药性强,OXA-23型基因可能是A.baumannii对碳青霉烯酶类抗菌药物耐药的主要原因,我院发现我国少见OXA-24基因或许为区域性流行,携带多种耐药基因是导致A.baumannii对多种常用抗菌药物耐药的重要原因。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.