慢性化脓性中耳炎的病原菌及耐药性分析

目的研究慢性化脓中耳炎主要病原菌的种类及对抗生素的耐药情况。方法对162例慢性化脓 性中耳炎患者的耳分泌物进行细菌培养,对分离的病原菌进行药物敏感试验。结果分离出病原菌183株, 以金黄色葡萄球菌(63株)和铜绿假单胞菌(57株)为主,金黄色葡萄球菌对青霉素和红霉素的耐药性率分别 为100％和71.4％,对氨基糖甙类、氟喹诺酮类和头孢类抗生素敏感性较高;铜绿假单胞茵对氨苄西林、头孢 类抗生素耐药率较高,对氟喹诺酮类、氨基糖甙类抗生素敏感。结论金黄色葡萄球菌和铜绿假单胞菌是济 南地区慢性化脓性中耳炎的主要致病菌,二者对青霉素类抗生素有较高的耐药率。临床医师不能仅凭经验, 而应根据细菌培养和药敏结果用药,以防耐药菌株产生。     

2个耳聋家系中常见耳聋基因的基因型与表型相关性分析

目的:检测耳聋家系中耳聋患者的基因型,结合听力损失程度,分析常见耳聋基因的基因型与表型之间的关系。方法:选取2个耳聋小家系,检测4个耳聋基因的9个位点的突变,同时对家系成员进行纯音测听检查。结果:LGR-1家系中,Ⅰ:1、Ⅰ:2、Ⅰ:4和Ⅱ:5基因型为单杂合突变,表型为轻中度聋;Ⅱ:3和Ⅱ:4基因型为双杂合突变,Ⅱ:2和Ⅲ:1基因型为复合杂合突变,表型均为重度或极重度聋。HXL-2家系中,Ⅰ:1和Ⅰ:2基因型为单杂合突变,表型为轻中度聋;Ⅱ:2、Ⅱ:3、Ⅱ:5和Ⅲ:2基因型为单杂合突变,但表型为重度或极重度聋;Ⅱ:1基因型为双杂合突变,表型为极重度聋;Ⅲ:1基因型为单杂合突变,左耳为中度聋,右耳为中重度聋。结论:单杂合突变患者表型多为轻度或中度聋,而基因型为单杂合突变的重度或极重度聋患者可能存在另外一个没有检测到的突变;基因型为双杂合突变或复合杂合突变的患者,表型多为重度或极重度聋;基因型与听力表型之间存在一定的正相关性。     

慢性化脓性中耳炎病原菌分析及临床意义

目的分析慢性化脓性中耳炎患者病原菌群分布及对抗生素的耐药性，指导临床用药。方法对收治的163例慢性化脓性中耳炎患者中耳分泌物进行细菌及真菌分离培养和药物敏感试验。结果163例患者中，有136例检出病原菌，检出率为83.4％。以金黄色葡萄球菌(42株)、铜绿假单胞菌(29株)和肺炎克雷伯菌（22株）为主,真菌6株。结论慢性化脓性中耳炎患者感染病原菌中以金黄色葡萄球菌多见，不同病原菌对药物的耐药性不同，临床使用抗生素时应行细菌培养及药物敏感试验，使用敏感抗生素治疗。     

阻塞性睡眠呼吸暂停低通气综合征患者CYP3A4基因多态性的检测

目的探讨阻塞性睡眠呼吸暂停低通气综合征(OSAHS)CYP3A4基因的多态性,了解其对芬太尼类药物的敏感性及不同基因型的人群分布特征,指导临床个体化用药。方法对50例OSAHS患者进行CYP3A4基因的多态性检测,采静脉血,通过DNA抽提-PCR扩增-焦磷酸测序方法检测CYP3A4基因。结果野生纯合型型34例(68%),突变杂合型15例(30%),突变纯合型型1例(2%)。结论OSAHS患者中约2%为AA型,该型对芬太尼类药物极其敏感,术后有易发生窒息的风险,应高度警惕芬太尼类药物呼吸抑制的潜在风险。     

慢性化脓性中耳炎病原菌及耐药性分析

目的研究慢性化脓性中耳炎的主要病原菌种类及对抗生寨的耐药情况。方法对132例慢性化脓性中耳炎患者的脓性分泌物进行细菌培养并对分离的病原菌进行药物敏感试验。结果分离出病原菌152株,以金黄色葡萄球菌（49株）和铜绿假单胞菌（25株）为主,真菌5株。金黄色葡萄球菌对喹诺酮类敏感,敏感率为79．6％。铜绿假单胞菌对头孢他啶和喹诺酮类敏感,敏感率分别为84％和64％。结论金黄色葡萄球菌和铜绿假单胞菌是聊城地区慢性化脓性中耳炎主要致病菌,二者均对喹诺酮类敏感,铜绿假单胞菌亦对三代头孢类的头孢他啶敏感。临床医师在对慢性化脓性中耳炎患者治疗时应根据细菌培养和药敏结果用药,准确有效的治疗,以防耐药菌株的发生。真菌在慢性化脓性中耳炎致病菌中不容忽视。     

一个非综合征型聋家系的遗传学分析及产前诊断

目的 对一个遗传性聋家系进行致病基因鉴定、遗传咨询和产前诊断。方法 运用目标区域捕获测序检测一个非综合征型聋家系遗传学病因,对检出的致病突变进行Sanger测序验证,结合STR检测技术对该家系行产前诊断。结果 一个家系两代人(Ⅰ:2、Ⅱ:2、Ⅱ:3)均为遗传性聋,但病因不同,先证者(Ⅰ:2)是SLC26A4基因C.919-2A>G纯合突变导致,两个异卵双胞胎女儿(Ⅱ:2、Ⅱ:3)耳聋病因是MYO15A基因c.5062＿5063delCT/c.7396-1G>A复合杂合突变。先证者孕期胎儿产前诊断结果显示耳聋风险低,出生后复查与产前诊断一致,并顺利通过新生儿听力筛查。结论 本研究明确了该非综合征型聋家系的基因型,首次明确了MYO15A基因c.5062＿5063delCT突变为致病性变异,拓展了MYO15A基因致病突变谱。     

顽固性中耳炎的微生物学特性

呼吸道病原菌对抗生素耐药日益加重,使急性中耳炎(acuteotitismedia,AOM)的控制亦变得困难。该研究报道作者近来对婴儿和儿童顽固性急性中耳炎中耳分离得到的病原体的评价。从1996年6月至1997年12月间患双侧AOM使用至少一周口服抗生素无效的儿童,经鼓膜穿刺抽吸中耳渗出液加以培养。46例患儿中有34例发现病原菌,分离得到43株病菌,发现1种病菌者29例,2种者7例。其中肺炎链球菌16株,流感嗜血杆菌无分类B型12株,卡他莫拉氏菌5株,化脓性链球菌5株,金黄色葡萄球菌3株以及胨链球菌2株。27株分离菌(63%)对抗生素有耐药性,耐阿莫西林以流感嗜血杆…     

呼吸喹诺酮类药物在耳鼻咽喉头颈外科中的应用进展

自1962年世界上第一个喹诺酮类药物萘啶酸被美国Lesher等[1]研发至今,喹诺酮类药物历经40余年的发展逐渐成为临床上接纳度较高的抗生素类药物之一.新一代的呼吸喹诺酮类药物近年来研究发展较为迅速,在耳鼻咽喉头颈外科领域的应用及研究前景受到了极大关注.     

莫西沙星在急性上呼吸道感染中的应用

莫西沙星(moxifloxacin)是第4代氟喹诺酮类抗菌药的代表药物。自1999年上市后已逐渐应用于临床,与其他喹诺酮类药物相比具有广谱、高效、低毒等特点。在国内主要用于重症下呼吸道感染和泌尿系统感染方面,在上呼吸道感染方面应用经验还不是很丰富。     

Waardenburg综合征Ⅱ型中国家系MITF基因突变分析

目的 探讨Waardenburg综合征Ⅱ型中国家系的临床和分子遗传学特征。方法收集两个Waardenburg综合征Ⅱ型中国家系详细的临床资料并绘制家系圈谱，签署知情同意书获取血样。聚合酶链反应扩增MITF基因编码区的全部外显子，在ABI3100自动测序仪上进行正反向测序，利用GeneTool软件及遗传学网站的信息分析数据。结果Waardenburg综合征Ⅱ型临床表现变异较大，不是所有的患者均满足Waardenburg综合征Ⅱ型的诊断标准，眼睛色素分布异常（蓝虹膜）和正常的内眦间距是临床上最常见的表型特征。MITF基因第1，7号外显子在两个隶系中分别检测到一个错义突变和一个缺失突变，50例正常对照组均未检测到这两种突变。结论本文对两个Waardenburg综合征Ⅱ型家系做出分子水平的基因诊断，其详细的临床资料有助于对该综合征的进一步认识；新的基因突变位点不仅丰富了人类基因突变数据库，而且为更好地了解MITF基因的功能提供了新的线索。     

Nasopharyngeal carriage of drug-resistant Streptococcus pneumoniae in children with acute otitis media evaluated by polymerase chain reaction-based genotyping of penicillin-binding proteins

A polymerase chain reaction (PCR)-based genotyping of the penicillin-binding protein (PBP) genes pbp1a, pbp2x and pbp2b was used to characterize Streptococcus pneumoniae isolated from the nasopharynx of children with acute otitis media (AOM). Mutations were observed in pbp1a, pbp2x and pbp2b genes in 36.5% of the strains. Decreased susceptibility to beta-lactam antibiotics was closely associated with the frequency of mutations in the three PBP genes. Of penicillin-intermediately-resistant S. pneumoniae strains, 54.5% appeared to be genetically similar to penicillin-resistant S. pneumoniae strains. Of penicillin-susceptible S. pneumoniae strains, 33.3% had mutations in the pbp2x gene and showed relatively high MICs to cephalosporins. Strains with mutations in the three PBP genes were often isolated from children < or = 2 years old. Evaluation of mutations in PBP genes using PCR will prove useful for studying the epidemiology of antibiotic resistance.     

Increase of macrolide-resistant Streptococcus pneumoniae-expressing mefE or ermB gene in the nasopharynx among children with otitis media

OBJECTIVE: To evaluate prevalence of macrolide resistant strains and the genotypes of the resistance among Streptococcus pneumoniae isolated from the nasopharynx of children with otitis media. STUDY DESIGN: Retrospective review. METHODS: A total of 858 S. pneumoniae isolates were collected from the nasopharynx of pediatric patients with acute otitis media at the clinics of Otolaryngology-Head and Neck Surgery, Wakayama Medical University Hospital and six affiliated hospitals in Wakayama prefecture between January 1998 and December 2002. The antibiotic susceptibility patterns were analyzed for penicillin, erythromycin, and clindamycin according to the National Committee for Clinical Laboratory Standards. Macrolide resistance genes of mefE and ermB were determined by polymerase chain reaction of all S. pneumoniae. RESULTS: Of 858 clinical isolates, 259 (30.1%) were strains without ermB or mefE gene, 279 (32.5%) carrying mefE, 292 (34.0%) carrying ermB, and 28 (3.4%) carrying both genes. There was a strong correlation between phenotypes and the presence of macrolide resistance genes. The macrolide resistance genes were especially frequently identified among penicillin-resistant S. pneumoniae. Strains carrying ermB gene gradually increased from 25% in 1998 to 45% in 2002, with a concurrent decrease in strains carrying mefE from 36% in 1998 to 1999 to 19% in 2002. Strains having mefE were frequently identified among children younger than 2 years old. The current finding suggested that high-level ermB-mediated macrolide resistance in S. pneumoniae is increasing at an alarming rate in pediatric patients with otitis media, especially among young children. Physicians should pay close attention to such macrolide-resistant bacterial pathogens in the antimicrobial treatment of pediatric patients with otitis media.     

Unilateral auditory neuropathy spectrum disorder

A polymerase chain reaction (PCR)-based genotyping of the penicillin-binding protein (PBP) genes pbp1a , pbp2x and pbp2b was used to characterize Streptococcus pneumoniae isolated from the nasopharynx of children with acute otitis media (AOM). Mutations were observed in pbp1a , pbp2x and pbp2b genes in 36.5% of the strains. Decreased susceptibility to &;#103 -lactam antibiotics was closely associated with the frequency of mutations in the three PBP genes. Of penicillin-intermediately-resistant S. pneumoniae strains, 54.5% appeared to be genetically similar to penicillin-resistant S. pneumoniae strains. Of penicillin-susceptible S. pneumoniae strains, 33.3% had mutations in the pbp2x gene and showed relatively high MICs to cephalosporins. Strains with mutations in the three PBP genes were often isolated from children &;#104 2 years old. Evaluation of mutations in PBP genes using PCR will prove useful for studying the epidemiology of antibiotic resistance.     

儿童慢性化脓性中耳炎病原菌分布及耐药性分析

目的探讨儿童慢性化脓性中耳炎病原菌分布及耐药性,为临床合理用药提供参考。方法以2015年1月至2019年12月郑州大学附属儿童医院耳鼻喉科收治的儿童慢性化脓性中耳炎患者401例(418耳)为研究对象,男163例,女238例,年龄2月21天~15岁,平均2.41±2.31岁。取中耳分泌物行细菌培养及药敏试验,采用WHONET 5.6软件进行药物敏感性分析。结果418耳慢性化脓性中耳炎中共391耳检出病原菌,排除取材时污染的7耳,共384耳,病原菌检出率为91.87%(384/418),其中细菌感染372耳,真菌感染12耳,革兰阳性菌检出率较高,为329株(85.68%);主要检出菌株依次为金黄色葡萄球菌168(43.75%)、肺炎链球菌106(27.60%)、表皮葡萄球菌26(6.77%)、铜绿假单胞菌19(4.95%)、嗜血杆菌13(3.39%)、真菌12(3.13%)株。常见致病菌对抗菌药物的敏感性因菌种而异,金黄色葡萄球菌对青霉素G、红霉素耐药率均>85%,肺炎链球菌对克林霉素、红霉素耐药率达100%,铜绿假单胞菌耐药率较高的依次为氨曲南和头孢吡肟。结论本组慢性化脓性中耳炎患儿的主要致病菌为金黄色葡萄球菌、肺炎链球菌、铜绿假单胞菌,临床上应根据致病菌的药敏试验结果合理选择抗菌药物,以确保患儿用药合理、安全、有效。     

High prevalence of Streptococcus pneumoniae with mutations in pbp1a, pbp2x, and pbp2b genes of penicillin-binding proteins in the nasopharynx in children in Japan

OBJECTIVE: To evaluate the resistances of Streptococcus pneumoniae to beta-lactams developed by stepwise alterations in high-molecular-weight penicillin-binding proteins (PBPs) with a reduced binding affinity of beta-lactams. Among the numerous mutations in pbp genes that alter the affinity for beta-lactams, the decreased affinity of PBP1A, 2X and 2B is especially important in the development of resistances to beta-lactams. STUDY DESIGN: Retrospective review. METHODS: In this study, we investigated the mutations in pbp1a, pbp2x, and pbp2b genes evaluated by polymerase chain reaction (PCR) in 866 pneumococcal isolates collected from the nasopharynx of Japanese children with acute otitis media. RESULTS: 210 strains (24.3%) exhibited no mutations in the three pbp genes. 333 strains (38.5%) had mutations in the three pbp genes, 78 (9.0%) in two pbp genes, whereas 245 (28.3%) displayed mutations in only one pbp gene. Among the 656 strains with mutations in pbp genes, 620 (94.5%) strains had mutations in pbp2x. The annual prevalence of antimicrobial-resistant S. pneumoniae showed a gradual increase in strains with mutations in the three pbp genes and a parallel decrease in strains without mutations. CONCLUSIONS: PCR-based genotyping can characterize the antimicrobial resistances in pneumococci along with minimal inhibitory concentrations (MICs). Physicians should pay attention to the recent increase in antimicrobial-resistant S. pneumoniae when treating pediatric acute otitis media.     

慢性中耳炎细菌生物膜形成与细菌培养结果的相关性分析

目的：探讨慢性中耳炎细菌生物膜形成与细菌培养之间的相关性。方法：对32例慢性化脓性中耳炎及中耳胆脂瘤患者术中取得的样本进行扫描电镜检查，并对中耳分泌物行细菌培养；分析慢性中耳炎细菌生物膜形成与细菌培养结果之间的关系。结果：慢性化脓性中耳炎（活动期）与中耳胆脂瘤细菌生物膜形成率分别为87．5％、81．3％（P〉0．05）。慢性中耳炎扫描电镜细菌生物膜结果与细菌培养结果之间比较，灵敏度为70．37％，特异度为60.00％，误诊率为40．00％，漏诊率为29．63％，阳性预测值为90．46％，阴性预测值为27．27％，正确率为68．75％，约登指数为30．37％，Pearson相关系数为0．232（P〉0．05）。结论：慢性化脓性中耳炎（活动期）与中耳胆脂瘤均有较高的生物膜形成率，但中耳分泌物常规细菌培养结果不能反应慢性中耳炎患者细菌生物膜的形成情况，需要探索更为可靠的细菌学实验方法来准确反映慢性中耳炎的感染情况。     

Molecular analysis of pathogens of upper respiratory tract infections in children--a study of nasopharyngeal S. pneumoniae and PBP genes in acute otitis media

We have recently been confronted with refractory upper respiratory infections with an increasing prevalence of penicillin (Pc)-resistant S. pneumoniae. There has been a broad consensus that acute otitis media (AOM) is caused by migration of pathogens from nasopharynx and proliferation in the middle ear space, and thus it is, very important to study the bacterial environment in the nasopharynx as the source of middle ear infections. Eighty pneumococcal isolates from the nasopharynx of children with acute otitis media were evaluated by polymerase chain reaction (PCR) for mutation of Pc-binding protein (PBP) genes. The results showed mutation of all three PBP genes, pbp 1a, pbp 2x, and pbp 2b, in 30% of the isolates, while 74% were found to possess various PBP gene mutations, mostly in one-year-old children. Of the 46 isolates whose minimum inhibitory concentration (MIC) of Pc was < or = 0.06 microgram/mL, 43% were found to possess a pbp 2x mutaion, which affects cefem resistance. We genotyped each pneumococcal isolate from the nasopharynx of children with recurrent AOM by pulsed-field gel electrophoresis (PFGE). In 9 of 11 pairs (82%) of consecutive AOM episodes, the nasopharyngeal isolate in the second episode was different. In addition, discrimination of each isolate based upon the mutation profile of its PBP genes in 8 pairs (72%) of consecutive AOM episodes showed that the isolates were different, and there was little difference between the results of PBP gene mutation and PFGE analysis. These findings suggest that most nasopharyngeal isolates from children with AOM possess PBP mutations and that children with increased numbers of drug-resistant bacteria in their nasopharynx during AOM has been colonized or recolonized by different strains during each episode. We therefore emphasize that clinicians should assess the antibiotic susceptibility of nasopharyngeal isolates from children during each episode. PBP gene mutation analysis of S. pneumoniae is useful not only in providing valuable information on the antibiotic susceptibility of each strain but for assessing changes in causative strains in the sequential episodes of pneumococcal infection.