儿童感染性腹泻是婴幼儿时期的常见病, 主要是由于婴幼儿时期消化系统发育还不完善, 并且机体的防御功能较差, 该病对儿童的健康危害严重, 治疗不及时或不恰当可导致死亡, 故加强对其的防治具有重要意义。
为明确青岛市儿童感染性腹泻的病原学分布特点及耐药性情况, 本研究对2013年青岛儿童医院肠道门诊就诊的急性感染性腹泻患儿粪便标作者单位：266012 青岛市妇女儿童医院检验科(薛爱国 丁伟), 呼吸科( 李海燕), 中心注射室(宋冬)通讯作者：丁伟婴幼儿腹泻病原菌分布特点及药敏分析薛爱国 李海燕 宋冬 丁伟【摘要】 目的 研究青岛市2013年儿童感染性腹泻的病原菌的分布、流行病学特征及对抗菌药物的敏感情况, 为临床预防以及合理用药提供参考依据。
方法 对2013年1月~2013年12月临床初步诊断为感染性腹泻患儿的新鲜粪便标本进行细菌分离培养和血清分型鉴定, 采用纸片扩散法进行药敏试验, 并以免疫层析双抗体夹心法定性检测A 群轮状病毒抗原。
结果 526份粪便培养共检出病原菌6种80株, 检出率为15.2%(80/526)。
检出沙门菌38株(47.5%), 志贺菌8株(10.0%), 致泻性大肠杆菌8株(10.0%), 金黄色葡萄球菌4株(5.0%), 铜绿假单胞菌19株(23.7%), 白假丝酵母菌3株(3.75%)。
8株志贺菌7株为福氏志贺菌, 1株鲍氏志贺菌。
沙门菌和志贺菌的耐药率分别为氨苄西林(68.7%、76.9%)、复方新诺明(21.1%、38.5%)、环丙沙星(15.8%、15.4%)、头孢曲松(6.25%、15.3%)。
526份标本中A 群轮状病毒阳性107份。
结论 2013年青岛市儿童细菌性腹泻的病原菌以沙门菌为主, 易感人群以2岁以下婴幼儿为主；而金黄色葡萄球菌感染以1岁以下婴儿为主；铜绿假单胞菌和白假丝酵母菌引起的腹泻主要是由于长期应用广谱抗生素使肠道菌群失调所导致；A 群轮状病毒是引起婴幼儿秋冬季腹泻的主要病原。
应规范临床用药, 加强细菌耐药监测。
【关键词】 儿童；感染性腹泻；病原学；耐药性；轮状病毒 Distribution characteristics of pathogens associated with infectious diarrhea in children from Qingdao district in 2012 and its sensitivity analysis XUE Ai-guo, LI Hai-yan, SONG Dong, et al. Department of Clinical Laboratory, Qingdao Women and Children Hospital, Qingdao 266012, China【Abstract 】 Objective To investigate the distribution of pathogens causing infectious diarrhea in children from Qingdao district in 2013 and analyze epidemiological characteristics and corresponding drug-sensitivity, in order to guide clinical prevention and treatment. Methods Pathogens were isolated and identified from fresh faeces of children preliminarily diagnosed with infectious diarrhea from January 2013 to December 2013. Drug-sensitivity testing was performed using paper disc diffusion method, and rotaviruses of A group were identified qualitatively with double antibody sandwich method to detect viral antigen. Results 80 strains of pathogens belonging to six species were isolated from 526 samples of faeces with the detection rate of 15.2% (80／526), including 38 strains of Salmonella (47.5%), 8 strains of Shigella (10.0%), 8 strains of diarrheogenic Escherichia coli (10.0%), 4 strains of Staphylococcus aureus (5.0%), 19 strains of Pseudomonas aeruginosa (23.7%) and 3 strains of Candidid albicans (3.75%). 7 strains of Shigella belonged to S. flexneri, 1 strain of Shigella belonged to S. boydii. Drug-sensitivity testing showed that for Salmonella and Shigella, the rates of drug resistance are as follows, to ampicillin, 60.5% and 87.5%, to SMZ-TMP, 21.1% and 37.5%, to Ciprofloxacin, 15.8% and 25%, to Ceftriaxone, 21.1% and 25%, respectively. A total of 107 samples were positive for rotavirus of group A. Conclusion The main pathogens causing bacterial diarrhea in children from Qingdao distrinct in 2012 are Shigella, mainly infecting infants below two years old, while Staphylococcus aureus prefer to infect infants below one year old. And bacterial diarrhea caused by Pseudomonas aeruginosa and Candidid albicans are induced mostly by enteric dysbacteriosis owing to long-term use of broad-spectrum antibiotics. Rotavirus of group A is the most common pathogen causing diarrhea of infants in autumn and winter. Therefore, clinical usage of antibotics should be regulated towards standardization and drug resistance should be monitored regularly.【Key words 】 Children; Infectious diarrhea; Pathogens; Drug-resistance; Rotavirus 本的细菌培养和药敏结果进行分析, 以期为临床的预防治疗提供依据。
1 资料与方法1. 1 一般资料 采集2013年1月~2013年12月本院肠道门诊中粪便常规镜检显示WBC ≥10/HPF、RBC ≥0/HPF, 临床初步诊断为急性腹泻病患儿的新鲜粪便标本526份, 置Carry-Blair 运送培养基, 立即做细菌培养, 检测沙门菌、志贺菌、致泻性大肠杆菌和金黄色葡萄球菌等；同时做快速A 群轮状病毒(RV)抗原检测。
1. 2 试剂来源 菌株鉴定使用法国梅里埃API鉴定试条；沙门菌、志贺菌、致泻性大肠杆菌诊断血清由宁波天润生物药业有限公司提供；药敏纸片使用英国Oxoid公司产品, 药敏培养基使用英国Oxoid生产的MH琼脂；标准菌株大肠埃希菌ATCC25922、金黄色葡萄球菌ATCC25923作为质控菌株, 由卫生部检验中心提供。
A群RV抗原检测试剂盒由北京万泰生物药业股份有限公司提供。
1. 3 细菌分离鉴定及药敏试验 细菌培养按《全国临床检验操作规程》操作。
粪便标本分别接种于血平板、麦康凯(MAC) 、SS平板(沙门菌和志贺菌琼脂平板)、35℃培养18~24 h后观察, 可疑菌落接种克氏双糖铁琼脂(KIA), 35℃培养过夜后, 进一步做血清分型鉴定。
药敏试验使用纸片扩散法, 结果按临床实验室标准化委员会(CLSI)制订的标准进行判断。
1. 4 A群RV抗原检测 采用免疫层析双抗体夹心法(胶体金法)快速定性检测。
2 结果2. 1 病原菌检出率及构成 526份粪便培养标本共检出病原菌6种80株, 检出率15.2%(80/526)。
病原菌检出率及构成比见表1。
表1 80株腹泻病原菌检出率及构成比菌种菌株数(n)检出率(%)构成比(%)沙门菌387.2247.5志贺菌8 1.5210致病性大肠杆菌8 1.5210铜绿假单胞菌19 3.6123.7金黄色葡萄球菌40.76 5.0白假丝酵母菌30.57 3.75合计8015.2100.0 2. 2 38株沙门菌阳性检出率 检出非伤寒沙门菌O4群18株, 占沙门菌检出总株数的47.3%；沙门菌O9群17株, 占44.7%；O7群、O8群、O12群各1株, 占2.6%。
2. 3 8株志贺菌阳性检出率 检出B群(福氏志贺菌)7株, 鲍氏志贺菌1株。
2. 4 药敏结果 沙门菌和志贺菌的药敏结果见表2。
表2 沙门菌和志贺菌的药敏情况抗生素沙门菌(n=32株)志贺菌(n=13株)敏感(S)耐药(R)敏感 (S)耐药(R)氨苄西林31.368.723.176.9头孢唑啉62.537.592.37.7头孢他啶93.75 6.2592.37.7头孢曲松93.75 6.2584.715.3头孢吡肟10001000头孢替坦90.69.486.213.8复方新诺明78.921.161.538.5庆大霉素59.440.684.615.4妥布霉素84.415.684.615.4阿米卡星62.537.584.615.4环丙沙星84.215.884.615.4亚胺培南10001000厄他培南10001000哌拉西林/他唑巴坦100010002. 5 A群RV感染 在526份临床初步诊断为感染性腹泻病患儿的粪便标本中, A群RV抗原检测阳性107份, 占20.3%。