本文主要研究内容
作者田二杰（2019）在《鸡源大肠杆菌对达氟沙星和安普霉素的耐药判定标准研究》一文中研究指出：对于禽类,一些大肠杆菌能够通过定植在肠道导致肠内感染,同时也能转移到肠外导致肠外感染,主要感染3～7周龄的雏鸡,临床表现为心包炎、肝周炎、腹膜炎、眼炎等症状,给养鸡业带来了巨大的经济损失。达氟沙星(氟喹诺酮类)和安普霉素(氨基糖苷类)是两种兽医专用广谱抗生素,常用于预防和治疗鸡、猪等动物的革兰氏阴性菌(如大肠杆菌)引起的感染。目前由于抗生素的长期不规范使用,大肠杆菌的耐药性问题愈发严重。然而国内外尚未建立这两种药物对大肠杆菌的耐药判定标准。本研究旨在通过建立达氟沙星和安普霉素的耐药判定标准,为规范这两种药物在我国兽医临床上的应用提供理论依据。(1)达氟沙星和安普霉素对大肠杆菌的野生型临界值(COWT)的测定本研究从全国各大规模化养鸡场采集并分离鉴定了 1412株鸡源大肠杆菌,采用微量肉汤稀释法测得达氟沙星和安普霉素的最小抑菌浓度(MIC)。达氟沙星在0.125～64 μg/mL浓度范围内的MIC呈双峰分布,峰值分别为0.5μg/mL和8 μg/mL,其中MIC为8μg/mL时细菌有215株,占15.7%,分布最多。达氟沙星对鸡大肠杆菌的MIC50为4μg/mL,MIC90为64μg/mL。而安普霉素在2～256 μg/mL浓度范围中呈单峰分布,峰值为MIC=8μg/mL,此处细菌有828株,占60.7%。安普霉素对大肠杆菌的MIC50为8μg/mL,MIC90为16μg/mL。将两种药物的MIC值转换为Log2MIC,进而采用非线性回归分析、NORMINV和NORMDIST方法建立达氟沙星和安普霉素的COWT分别为4 μg/mL和16 μg/mL。(2)大肠杆菌对达氟沙星和安普霉素的耐药特点分析选择不同MIC值的大肠杆菌,用聚合酶链式反应(PCR)法检测耐药基因与MIC的相关性。结果表明大肠杆菌中达氟沙星耐药基因oqxAB、aac(Ib-cr)、qnrAqnrB、qnrS和qepA基因的阳性率在一定程度上随着MIC值的升高而增大,qnrC和qnrD阳性率为0。同样,安普霉素耐药基因aac(3)-Ⅳ和npmA的阳性率也具有MIC值依赖性。采用微量肉汤稀释法进一步筛选到24株达氟沙星高度耐药菌和16株安普霉素高度耐药菌,并使用纸片扩散法检测这两类菌株对17种抗革兰氏阴性菌常用抗生素的敏感性,结果表明这两类菌株均具有多重耐药性,最低10耐,最高达15～16耐。同时还发现所有的达氟沙星耐药菌株对阿莫西林、氨苄西林、萘啶酸、恩诺沙星、环丙沙星、多西环素、四环素和甲氧苄啶均表现出100%耐药,而对阿米卡星和磷霉素相对较敏感。所有的安普霉素耐药菌株对阿莫西林、氨苄西林、庆大霉素、强力霉素、四环素、甲氧苄啶和氟苯尼考均表现出100%耐药。所有安普霉素耐药菌株都对庆大霉素耐药,而对阿米卡星和大观霉素相对较敏感。(3)达氟沙星对大肠杆菌的药效学临界值(COPD)的建立构建大肠杆菌感染鸡动物模型,单次口服给予达氟沙星5 mg/kg后,分别收集不同时间点健康组和感染组鸡的血样和4种肠段(十二指肠、空肠、回肠和盲肠)的肠液,并用高效液相色谱荧光法检测药物浓度。4种肠段的群体药物代谢动力学特点符合二室模型。健康组鸡从十二指肠、空肠到回肠的吸收速率(Ka)、清除率(CL)和分布体积(V)依次下降。而在感染组鸡的肠道中,空肠中Ka和V值最高。与健康组相比,感染组鸡从十二指肠、空肠到回肠,达氟沙星的CL依次降低,且均低于健康组鸡的相应肠段,十二指肠和回肠的Ka显著降低,空肠和回肠中的V增大。健康组和感染组鸡回肠中的F均较高。大肠杆菌078在MH肉汤、回肠液和血浆中的生长曲线的趋势一致。微量肉汤稀释法检测达氟沙星在肉汤、血浆和4种肠液中对大肠杆菌078的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果显示,达氟沙星在MH肉汤和血浆中的MIC均为0.016μg/mL,而在4种肠液中MIC值均比血浆中和MH肉汤中的大,其中空肠液中的MIC值最大(6.4 μg/mL),而在回肠液中的MIC值最小(0.64 μg/mL)。采用平板倾倒计数法,绘制达氟沙星对大肠杆菌078的体外和半体内(回肠和十二指肠)杀菌曲线。达氟沙星的体外和半体内杀菌作用均表现出浓度依赖性,因此可选择PK/PD参数AUC/MIC来制定COPD。综合血浆和4种肠段中达氟沙星的药动学(PK)和药效学(PD)特点,最终采用感染组回肠中的数据建立COPD。使用Winnonlin软件分析血浆和回肠的PK数据,得到达氟沙星在感染组鸡血浆中的平均达峰时间(Tmax)、达峰浓度(Cmax)和药时曲线下面积(AUC24)分别为 1 h,0.38 μg/mL 和 2.06 h·μg/mL;在回肠中的 Tmax、Cmax和 AUC 分别为 2.7 h,18.95μg/mL和261.78 h·μg/mL。进而用Winnonlin软件中Sigmoid Emax模型对回肠的PK/PD数据进行模拟,选择E=-3时AUC/MIC的值为药效学目标。计算得到达氟沙星在回肠中的药效学目标为421.45。最后使用Crystal ball软件对达氟沙星在回肠中的PK/PD数据进行蒙特卡洛模拟,得到不同MIC值下AUC/MIC达到药效学目标的概率,选择达标率大于90%的最大MIC为COPD。即回肠液中达氟沙星对大肠杆菌的COPD为0.54 μg/mL。(4)达氟沙星对大肠杆菌的临床临界值(COCL)的制定本研究中,达氟沙星对鸡的临床治疗试验包括感染组、感染治疗组和1个空白对照组,每组10只鸡。攻毒菌株是通过PCR和雏鸡攻毒实验筛选出的5个不同MIC值的大肠杆菌致病株。每一个感染组对应三个感染治疗组,治疗方案分别为5 mg/kg,2次/天,连续3天;10mg/kg,2次/天,连续3天;20mg/kg,1次/天,连续3天。治疗结果表明,3种治疗方案的治愈率无明显差异。因此,选择5 mg/kg的治疗结果来制定COCL。首先采用EUCAST推荐的WindoW法,得到达氟沙星对大肠杆菌的COCL范围为0.5～16 μg/mL。然后用CART分类树回归分析法分析得到,当POC等于90%时,COCL>2.25μg/mL。因此,推荐4μg/mL为达氟沙星对大肠杆菌的COCL。综上,本研究中达氟沙星的COWT=COcL=4μg/mL,COPD为0.54μg/mL。因此,达氟沙星的耐药判定标准为4 μg/mL。而安普霉素的COWT为16 μg/mL,其药效学和临床临界值还有待进一步研究和确定。
Abstract
dui yu qin lei ,yi xie da chang gan jun neng gou tong guo ding zhi zai chang dao dao zhi chang nei gan ran ,tong shi ye neng zhuai yi dao chang wai dao zhi chang wai gan ran ,zhu yao gan ran 3～7zhou ling de chu ji ,lin chuang biao xian wei xin bao yan 、gan zhou yan 、fu mo yan 、yan yan deng zheng zhuang ,gei yang ji ye dai lai le ju da de jing ji sun shi 。da fu sha xing (fu kui nuo tong lei )he an pu mei su (an ji tang gan lei )shi liang chong shou yi zhuan yong an pu kang sheng su ,chang yong yu yu fang he zhi liao ji 、zhu deng dong wu de ge lan shi yin xing jun (ru da chang gan jun )yin qi de gan ran 。mu qian you yu kang sheng su de chang ji bu gui fan shi yong ,da chang gan jun de nai yao xing wen ti yu fa yan chong 。ran er guo nei wai shang wei jian li zhe liang chong yao wu dui da chang gan jun de nai yao pan ding biao zhun 。ben yan jiu zhi zai tong guo jian li da fu sha xing he an pu mei su de nai yao pan ding biao zhun ,wei gui fan zhe liang chong yao wu zai wo guo shou yi lin chuang shang de ying yong di gong li lun yi ju 。(1)da fu sha xing he an pu mei su dui da chang gan jun de ye sheng xing lin jie zhi (COWT)de ce ding ben yan jiu cong quan guo ge da gui mo hua yang ji chang cai ji bing fen li jian ding le 1412zhu ji yuan da chang gan jun ,cai yong wei liang rou shang xi shi fa ce de da fu sha xing he an pu mei su de zui xiao yi jun nong du (MIC)。da fu sha xing zai 0.125～64 μg/mLnong du fan wei nei de MICcheng shuang feng fen bu ,feng zhi fen bie wei 0.5μg/mLhe 8 μg/mL,ji zhong MICwei 8μg/mLshi xi jun you 215zhu ,zhan 15.7%,fen bu zui duo 。da fu sha xing dui ji da chang gan jun de MIC50wei 4μg/mL,MIC90wei 64μg/mL。er an pu mei su zai 2～256 μg/mLnong du fan wei zhong cheng chan feng fen bu ,feng zhi wei MIC=8μg/mL,ci chu xi jun you 828zhu ,zhan 60.7%。an pu mei su dui da chang gan jun de MIC50wei 8μg/mL,MIC90wei 16μg/mL。jiang liang chong yao wu de MICzhi zhuai huan wei Log2MIC,jin er cai yong fei xian xing hui gui fen xi 、NORMINVhe NORMDISTfang fa jian li da fu sha xing he an pu mei su de COWTfen bie wei 4 μg/mLhe 16 μg/mL。(2)da chang gan jun dui da fu sha xing he an pu mei su de nai yao te dian fen xi shua ze bu tong MICzhi de da chang gan jun ,yong ju ge mei lian shi fan ying (PCR)fa jian ce nai yao ji yin yu MICde xiang guan xing 。jie guo biao ming da chang gan jun zhong da fu sha xing nai yao ji yin oqxAB、aac(Ib-cr)、qnrAqnrB、qnrShe qepAji yin de yang xing lv zai yi ding cheng du shang sui zhao MICzhi de sheng gao er zeng da ,qnrChe qnrDyang xing lv wei 0。tong yang ,an pu mei su nai yao ji yin aac(3)-Ⅳhe npmAde yang xing lv ye ju you MICzhi yi lai xing 。cai yong wei liang rou shang xi shi fa jin yi bu shai shua dao 24zhu da fu sha xing gao du nai yao jun he 16zhu an pu mei su gao du nai yao jun ,bing shi yong zhi pian kuo san fa jian ce zhe liang lei jun zhu dui 17chong kang ge lan shi yin xing jun chang yong kang sheng su de min gan xing ,jie guo biao ming zhe liang lei jun zhu jun ju you duo chong nai yao xing ,zui di 10nai ,zui gao da 15～16nai 。tong shi hai fa xian suo you de da fu sha xing nai yao jun zhu dui a mo xi lin 、an bian xi lin 、nai ding suan 、en nuo sha xing 、huan bing sha xing 、duo xi huan su 、si huan su he jia yang bian ding jun biao xian chu 100%nai yao ,er dui a mi ka xing he lin mei su xiang dui jiao min gan 。suo you de an pu mei su nai yao jun zhu dui a mo xi lin 、an bian xi lin 、qing da mei su 、jiang li mei su 、si huan su 、jia yang bian ding he fu ben ni kao jun biao xian chu 100%nai yao 。suo you an pu mei su nai yao jun zhu dou dui qing da mei su nai yao ,er dui a mi ka xing he da guan mei su xiang dui jiao min gan 。(3)da fu sha xing dui da chang gan jun de yao xiao xue lin jie zhi (COPD)de jian li gou jian da chang gan jun gan ran ji dong wu mo xing ,chan ci kou fu gei yu da fu sha xing 5 mg/kghou ,fen bie shou ji bu tong shi jian dian jian kang zu he gan ran zu ji de xie yang he 4chong chang duan (shi er zhi chang 、kong chang 、hui chang he mang chang )de chang ye ,bing yong gao xiao ye xiang se pu ying guang fa jian ce yao wu nong du 。4chong chang duan de qun ti yao wu dai xie dong li xue te dian fu ge er shi mo xing 。jian kang zu ji cong shi er zhi chang 、kong chang dao hui chang de xi shou su lv (Ka)、qing chu lv (CL)he fen bu ti ji (V)yi ci xia jiang 。er zai gan ran zu ji de chang dao zhong ,kong chang zhong Kahe Vzhi zui gao 。yu jian kang zu xiang bi ,gan ran zu ji cong shi er zhi chang 、kong chang dao hui chang ,da fu sha xing de CLyi ci jiang di ,ju jun di yu jian kang zu ji de xiang ying chang duan ,shi er zhi chang he hui chang de Kaxian zhe jiang di ,kong chang he hui chang zhong de Vzeng da 。jian kang zu he gan ran zu ji hui chang zhong de Fjun jiao gao 。da chang gan jun 078zai MHrou shang 、hui chang ye he xie jiang zhong de sheng chang qu xian de qu shi yi zhi 。wei liang rou shang xi shi fa jian ce da fu sha xing zai rou shang 、xie jiang he 4chong chang ye zhong dui da chang gan jun 078de zui xiao yi jun nong du (MIC)he zui xiao sha jun nong du (MBC)。jie guo xian shi ,da fu sha xing zai MHrou shang he xie jiang zhong de MICjun wei 0.016μg/mL,er zai 4chong chang ye zhong MICzhi jun bi xie jiang zhong he MHrou shang zhong de da ,ji zhong kong chang ye zhong de MICzhi zui da (6.4 μg/mL),er zai hui chang ye zhong de MICzhi zui xiao (0.64 μg/mL)。cai yong ping ban qing dao ji shu fa ,hui zhi da fu sha xing dui da chang gan jun 078de ti wai he ban ti nei (hui chang he shi er zhi chang )sha jun qu xian 。da fu sha xing de ti wai he ban ti nei sha jun zuo yong jun biao xian chu nong du yi lai xing ,yin ci ke shua ze PK/PDcan shu AUC/MIClai zhi ding COPD。zeng ge xie jiang he 4chong chang duan zhong da fu sha xing de yao dong xue (PK)he yao xiao xue (PD)te dian ,zui zhong cai yong gan ran zu hui chang zhong de shu ju jian li COPD。shi yong Winnonlinruan jian fen xi xie jiang he hui chang de PKshu ju ,de dao da fu sha xing zai gan ran zu ji xie jiang zhong de ping jun da feng shi jian (Tmax)、da feng nong du (Cmax)he yao shi qu xian xia mian ji (AUC24)fen bie wei 1 h,0.38 μg/mL he 2.06 h·μg/mL;zai hui chang zhong de Tmax、Cmaxhe AUC fen bie wei 2.7 h,18.95μg/mLhe 261.78 h·μg/mL。jin er yong Winnonlinruan jian zhong Sigmoid Emaxmo xing dui hui chang de PK/PDshu ju jin hang mo ni ,shua ze E=-3shi AUC/MICde zhi wei yao xiao xue mu biao 。ji suan de dao da fu sha xing zai hui chang zhong de yao xiao xue mu biao wei 421.45。zui hou shi yong Crystal ballruan jian dui da fu sha xing zai hui chang zhong de PK/PDshu ju jin hang meng te ka luo mo ni ,de dao bu tong MICzhi xia AUC/MICda dao yao xiao xue mu biao de gai lv ,shua ze da biao lv da yu 90%de zui da MICwei COPD。ji hui chang ye zhong da fu sha xing dui da chang gan jun de COPDwei 0.54 μg/mL。(4)da fu sha xing dui da chang gan jun de lin chuang lin jie zhi (COCL)de zhi ding ben yan jiu zhong ,da fu sha xing dui ji de lin chuang zhi liao shi yan bao gua gan ran zu 、gan ran zhi liao zu he 1ge kong bai dui zhao zu ,mei zu 10zhi ji 。gong du jun zhu shi tong guo PCRhe chu ji gong du shi yan shai shua chu de 5ge bu tong MICzhi de da chang gan jun zhi bing zhu 。mei yi ge gan ran zu dui ying san ge gan ran zhi liao zu ,zhi liao fang an fen bie wei 5 mg/kg,2ci /tian ,lian xu 3tian ;10mg/kg,2ci /tian ,lian xu 3tian ;20mg/kg,1ci /tian ,lian xu 3tian 。zhi liao jie guo biao ming ,3chong zhi liao fang an de zhi yu lv mo ming xian cha yi 。yin ci ,shua ze 5 mg/kgde zhi liao jie guo lai zhi ding COCL。shou xian cai yong EUCASTtui jian de WindoWfa ,de dao da fu sha xing dui da chang gan jun de COCLfan wei wei 0.5～16 μg/mL。ran hou yong CARTfen lei shu hui gui fen xi fa fen xi de dao ,dang POCdeng yu 90%shi ,COCL>2.25μg/mL。yin ci ,tui jian 4μg/mLwei da fu sha xing dui da chang gan jun de COCL。zeng shang ,ben yan jiu zhong da fu sha xing de COWT=COcL=4μg/mL,COPDwei 0.54μg/mL。yin ci ,da fu sha xing de nai yao pan ding biao zhun wei 4 μg/mL。er an pu mei su de COWTwei 16 μg/mL,ji yao xiao xue he lin chuang lin jie zhi hai you dai jin yi bu yan jiu he que ding 。
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论文作者分别是来自东北农业大学的田二杰，发表于刊物东北农业大学2019-08-27论文，是一篇关于大肠杆菌论文,达氟沙星论文,安普霉素论文,耐药判定标准论文,野生型临界值论文,东北农业大学2019-08-27论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自东北农业大学2019-08-27论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：大肠杆菌论文; 达氟沙星论文; 安普霉素论文; 耐药判定标准论文; 野生型临界值论文; 东北农业大学2019-08-27论文;