深圳市408份不同来源样本弯曲菌的检测及耐药特征分析

目的 分析深圳市不同来源样本弯曲菌的流行和抗生素敏感特征,与往年菌株的耐药特征进行比对,获得深圳地区弯曲菌的流行现状及耐药变化。方法 采用增强过滤法分离弯曲菌并应用琼脂稀释法进行抗生素敏感性分析。结果 408份不同来源样本中,鸡盲肠的弯曲菌阳性率最高,达72.0%(36/50),鸡肉的分离率次之,为49.3%(34/69)。腹泻病人弯曲菌分离率为10.7%(17/159)。牛肉、猪肉中弯曲菌分离率低,海鲜中分离到1株空肠弯曲菌,生菜未分离到弯曲菌。53株腹泻病人来源和鸡源空肠弯曲菌耐药率最高的是萘啶酸(94.3%)和环丙沙星(94.3%),其次是四环素(92.4%)、氟苯尼考(75.5%)。32株腹泻病人来源和鸡源结肠弯曲菌耐药率最高为萘啶酸、环丙沙星、四环素(100%)。73.6%(39/53)的空肠弯曲菌和100%(32/32)的结肠弯曲菌存在多重耐药性。与往年相比,鸡源空肠弯曲菌对大环内酯类抗生素的耐药增强,腹泻病人来源空肠弯曲菌对萘啶酸-环丙沙星-四环素-氟苯尼考多重耐药性增强。结论本地区首次在腹泻患者中分离到简明弯曲菌,其病原特征有待于进一步分析。深圳地区腹泻病人来源及鸡源空肠弯曲菌有耐药增强现象,应引起重视。     

鸡肉生产链空肠弯曲杆菌的耐药性研究

目的 了解鸡肉生产链(养殖场-屠宰场-市场)中空肠弯曲杆菌分离株的耐药情况。方法 针对成都市及雅安市养鸡场、屠宰场和市场中分离的180株空肠弯曲杆菌,采用微量肉汤稀释法,检测鸡肉生产链中空肠弯曲杆菌对8种抗菌药物的耐药情况;采用错配扩增突变分析PCR(MAMA PCR)技术,检测137株耐环丙沙星的空肠弯曲杆菌的gyrA基因第257位突变情况,对gyrA基因突变进行分析。结果 药敏试验表明,空肠弯曲杆菌分离株对环丙沙星、左氟沙星、四环素和克林霉素的耐药率较高,分别为94.44%、93.89%、91.67%和78.33%;对红霉素(20.00%)、链霉素(20.56%)、庆大霉素(17.78%)、氟苯尼考(12.22%)呈现不同的耐药率。从养殖场到市场,鸡肉生产链中空肠弯曲杆菌对上述8种抗菌药物的耐药率呈持平或上升趋势。MAMA PCR 结果表明,95.62%(131/137)的环丙沙星耐药空肠弯曲杆菌在gyrA基因第257位发生点突变,10株环丙沙星敏感菌株均没有检测出gyrA基因的突变;gyrA基因耐药决定区的测序结果表明,环丙沙星耐药菌株MJF31和MJF53都在第257位碱基发生了ACA→ATA(苏氨酸到异亮氨酸)的突变,而对环丙沙星敏感的菌株JF9和JF136均未发生该类突变。结论 鸡肉生产链中空肠弯曲杆菌耐药性比较普遍,对喹诺酮类药物耐药率较高,多重耐药菌株较突出,应加强对养殖场和食品源空肠弯曲杆菌耐药性的监测。     

福建地区鼠伤寒沙门菌喹诺酮耐药基因特征及MLVA分子分型

目的了解福建地区鼠伤寒沙门菌喹诺酮耐药状况及其分子分型机制。方法采用微量肉汤稀释法测定萘啶酸、环丙沙星和左氧氟沙星对83株鼠伤寒沙门菌的最小抑菌浓度(MIC);PCR扩增沙门菌的喹诺酮耐药区(QRDR)基因(gyrA,parC)和质粒介导的喹诺酮耐药基因(PMQR)(aac(6’)-Ib-cr,oqxA,oqxB);采用多位点可变数目串联重复序列分析(MLVA)比较菌株间的遗传相似性。结果 83株鼠伤寒沙门菌对萘啶酸、环丙沙星和左氧氟沙星的耐药率分别为27.7%(23/83),13.2%(11/83)和4.8%(4/83),QRDR基因突变率为25.3%(21/83)。21株基因突变株存在gyrA和parC两个突变类型,突变氨基酸类型分别为Ser83Tyr、Ser83Phe、Asp87Tyr和Asp87Asn,其中以第87位氨基酸突变为主,占总突变数的52.4%(11/21)。parC基因突变菌1株,为第80位氨基酸突变Ser80Arg,该菌株同时为gyrA:Ser83Phe突变。PMQR基因(aac(6’)-Ib-cr,oqxA,oqxB)在83株鼠伤寒沙门菌中的检出率分别为20.5%(17/83)、15.7%(13/83)和16.9%(14/83)。72株鼠伤寒沙门菌经MLVA分型后遗传关联度介于42.3%～100%之间,按照100%的相似度可分为18个型别,其中次优势型MT5菌株耐药基因与MLVA分型表现出一定的关联性。结论福建地区鼠伤寒沙门菌对喹诺酮类药物的耐药率不高,但环丙沙星和左氧氟沙星中度敏感菌中存在耐药基因突变,并具有一定的聚集性,需继续加强检测。     

福建地区鼠伤寒沙门菌喹诺酮耐药基因特征及MLVA分子分型北大核心CSCD

目的了解福建地区鼠伤寒沙门菌喹诺酮耐药状况及其分子分型机制。方法采用微量肉汤稀释法测定萘啶酸、环丙沙星和左氧氟沙星对83株鼠伤寒沙门菌的最小抑菌浓度(MIC);PCR扩增沙门菌的喹诺酮耐药区(QRDR)基因(gyrA,parC)和质粒介导的喹诺酮耐药基因(PMQR)(aac(6’)-Ib-cr,oqxA,oqxB);采用多位点可变数目串联重复序列分析(MLVA)比较菌株间的遗传相似性。结果 83株鼠伤寒沙门菌对萘啶酸、环丙沙星和左氧氟沙星的耐药率分别为27.7%(23/83),13.2%(11/83)和4.8%(4/83),QRDR基因突变率为25.3%(21/83)。21株基因突变株存在gyrA和parC两个突变类型,突变氨基酸类型分别为Ser83Tyr、Ser83Phe、Asp87Tyr和Asp87Asn,其中以第87位氨基酸突变为主,占总突变数的52.4%(11/21)。parC基因突变菌1株,为第80位氨基酸突变Ser80Arg,该菌株同时为gyrA:Ser83Phe突变。PMQR基因(aac(6’)-Ib-cr,oqxA,oqxB)在83株鼠伤寒沙门菌中的检出率分别为20.5%(17/83)、15.7%(13/83)和16.9%(14/83)。72株鼠伤寒沙门菌经MLVA分型后遗传关联度介于42.3%~100%之间,按照100%的相似度可分为18个型别,其中次优势型MT5菌株耐药基因与MLVA分型表现出一定的关联性。结论福建地区鼠伤寒沙门菌对喹诺酮类药物的耐药率不高,但环丙沙星和左氧氟沙星中度敏感菌中存在耐药基因突变,并具有一定的聚集性,需继续加强检测。     

宋内志贺菌对氟喹诺酮类药物敏感性降低的相关耐药基因研究

目的 研究宋内志贺菌对氟喹诺酮类抗菌药物敏感性降低的相关耐药基因的变化情况.方法 采用琼脂稀释法对131株宋内志贺菌进行药物敏感性检测,采用PCR法检测DNA旋转酶A亚单位(gyrA)、拓扑异构酶ⅣC亚单位(parC)基因的喹诺酮类药物耐药决定区(QRDR),并对PCR结果进行测序分析,同时用PCR法对质粒介导喹诺酮类耐药(PMQR)基因(qnr)和氨基糖苷乙酰转移酶变异基因aac(6’)-Ib-cr]进行筛选.结果 131株宋内志贺菌对萘啶酸的耐药率达100％,而对诺氟沙星、环丙沙星、左氧氟沙星全部敏感,对四环素、复方磺胺甲(噁)唑和氨苄西林的耐药率分别为93.9％、92.8％和93.2％.94％的萘啶酸耐药宋内志贺菌株对氟喹诺酮类药物敏感性出现下降.萘啶酸耐药菌株gyrA均发生单点83位丝氨酸→亮氨酸(Ser→ Leu)突变,但parC未发生突变;未检出qnr和aac(6')-Ib-cr基因.结论 萘啶酸耐药宋内志贺菌对氟喹诺酮类药物敏感性降低,其产生的主要机制为gyrA发生83位Ser→Leu替换.     

福氏志贺菌喹诺酮类耐药临床分离株gyrA和parC的基因突变

目的检测福氏志贺菌喹诺酮类耐药临床分离株DNA旋转酶gyrA和拓扑异构酶ⅣparC基因的突变情况，探讨GyrA和ParC氨基酸改变与喹诺酮类耐药的相关性。方法根据药敏实验结果选取47株福氏志贺菌临床分离菌，对其gyrA、parC喹诺酮耐药决定区（QRDR）进行聚合酶链反应（PCR）扩增和测序分析，并采用SAS（V8．2）软件分析GyrA和ParC改变和喹诺酮类耐药性的关联程度。结果44株喹诺酮类耐药菌在gyrA、parC基因QRDR均发生有义突变，gyrA83位密码子突变（TCG→TTG）最为常见．存在于43株耐药菌。混合模型分析结果显示GyrA83位、ParC80位氨基酸改变与萘啶酸的耐药性密切相关（P〈0．01，R^2=0．999），GyrA83、87位氨基酸改变与环丙沙星的耐药性密切相关（P〈0．05，R^2=0．872）。结论GyrA Ser83→Leu突变是导致福氏志贺菌临床株对喹诺酮类耐药的关键突变，此单位点突变即可介导福氏志贺菌对萘啶酸的耐药，但对环丙沙星耐药需同时存在GyrA和／或ParC多个位点突变或其他耐药机制。     

肉鸡源多重耐药空肠、结肠弯曲菌的耐药分子特征

目的了解肉鸡源空肠和结肠弯曲菌的耐药谱特征,检测多重耐药菌株Ⅰ类整合子/基因盒、gyrA基因突变位点、tetO基因、23S rRNA突变位点的分子特征。方法利用PCR检测弯曲菌Ⅰ类整合子/基因盒的存在情况;利用MAMA PCR技术检测弯曲菌gyrA基因第257位碱基的突变情况;针对弯曲菌23S rRNA的V区2075突变位点检测突变菌株。结果多重耐药菌株占分离株的94.5%。146株多重耐药空肠和结肠弯曲菌中Ⅰ类整合子检出率为98.6%,有78株菌株检出3种基因盒,1 000 bp+750 bp+500 bp+250 bp为主要谱型,所占比例为92.3%;有131株在gyrA喹诺酮类耐药决定区发生突变,突变率为92.9%。127株四环素耐药弯曲菌tetO基因的检出率为95.3%。81株红霉素耐药菌株中,23S rRNA的V区2075处突变发生率为96.3%。结论空肠和结肠弯曲菌分离株携带aadA2耐药基因盒,与氨基糖苷类药物的耐药性相关;gyrA基因突变、tetO基因的携带以及23S rRNA突变,与弯曲菌对喹诺酮、四环素和大环内酯类耐药密切相关。     

安徽地区腹泻患者感染肠致病大肠埃希氏菌的耐药特征

目的 对安徽地区腹泻患者粪便标本中肠致病大肠埃希氏菌(EPEC)进行检测,了解本地区EPEC的检出情况、耐药特征及耐药机制。方法 使用多重实时荧光定量PCR法对分离疑似菌株进行鉴定,微量肉汤稀释法对EPEC进行药物敏感性检测,使用双纸片扩散法检测ESBLs,采用多重PCR法检测ESBLs基因型;根据药敏结果,进行基因扩增和测序分析检测喹诺酮类耐药基因。结果 2013-2017年,本地区10家哨点医院共检出不典型EPEC 85株,典型EPEC和艾伯特埃希氏菌各1株;87株菌对红霉素、氨苄西林、甲氧苄啶/磺胺甲恶唑、萘啶酸和头孢噻肟耐药率分别为:100%、51.7%、40.2%、34.5%和20.7%,对亚胺培南、头孢他定、头孢西丁和环丙沙星耐药率分别为:0、3.4%、5.7%和6.9%。87株菌中检测出产ESBLs 18株,TEM阳性14株、CTX-M group 1阳性6株、CTX-M group 9阳性7株。喹诺酮类耐药机制有:药物靶位及编码基因gyrA和parC突变;质粒介导的qnrA、qnrB、qnrS和aac(6′)-Ib耐药基因阳性;药物外排泵qepA和oqxAB基因阳性。结论 EPEC是本地区重要的肠道致病菌之一,对青霉素、红霉素和萘啶酸耐药率较高,对亚胺培南、环丙沙星和阿奇霉素耐药率较低;ESBLs检出率较高,基因型以TEM、CTX-M group 1和CTX-M group 9为主,对喹诺酮类药物耐药机制主要与gyrA、parC突变和Qnr蛋白家族相关,少数与QepA 和OqxAB外排泵相关。     

宁波地区耐多药结核分枝杆菌喹诺酮耐药gyr基因突变研究

目的 为阐明宁波地区耐多药结核分枝杆菌(Multiple drug-resistant tuberculosis, MDR-TB)的gyr基因突变特征,深入研究MDR-TB对喹诺酮类药物耐药与gyr基因突变特征的关系。方法 采用1%比例法对MDR-TB进行氧氟沙星药敏检测实验,通过 DNA直接测序法分析MDR-TB的gyr基因突变情况。结果 120株MDR-TB临床分离株中有34株对喹诺酮耐药,总耐药率为28.33%(34/120)。34株耐喹诺酮菌株中,30株gyr基因发生突变,突变率为88.24%(30/34)。30株gyr基因发生突变的菌株中gyrA基因突变有29株,占96.67%(29/30),突变位点包括90、91和94位氨基酸;gyrB基因突变有2株,其中1株均合并gyrA基因突变,占6.67%(2/30),突变位点包括499和502位氨基酸。结论 宁波地区MDR-TB对喹诺酮类药物耐药形势较为严峻,gyrA基因突变与MDR-TB对喹诺酮类药物耐药相关。     

肉鸡屠宰加工生产链中弯曲菌污染状况及耐药性分析

目的 了解江苏某肉鸡屠宰加工厂不同环节弯曲菌的污染状况及耐药现状。方法 采集屠宰前肉鸡泄殖腔、不同环节鸡酮体以及环境等样品,运用无血弯曲杆菌选择性培养基(modified charcoal-cefoperazone-deoxycholate agar,CCDA)平板法进行分离纯化,PCR鉴定后,采用K-B法测定6大类14种抗生素耐药情况。PCR检测弯曲菌Ⅰ型整合子、四环素耐药基因tet(O)、氨基糖苷类耐药基因簇aadE-sat4-aphA-3以及弯曲菌gyrA基因第257位碱基的突变情况。结果 细菌分离结果显示170份样品分离到124株弯曲菌(72.95%)。分离株对链霉素、妥布霉素、卡那霉素、环丙沙星、氧氟沙星、萘啶酸、红霉素和四环素耐药率为100%,多重耐药现象普遍,多重耐药率高达100%,其优势耐药谱主要为CTX-CRO-cDA-K-NOR-S-AZM-CIP-TE-NA-E-LEV-ENR-TOB。分离株经PCR扩增未检测到Ⅰ型整合子,所有菌株在gyrA喹诺酮类耐药决定区发生了C-257-T点突变,tet(O)基因和aadE-sat4-aphA-3检出率分别为100%和90.23%。结论 江苏某肉鸡屠宰加工生产链中弯曲菌污染及耐药情况比较严重,gyrA基因突变、tet(O)基因及aadE-sat4-aphA-3耐药基因簇的携带,与弯曲菌对喹诺酮类、四环素类和氨基糖苷类耐药密切相关。     

Emergence and properties of fluoroquinolone resistant Salmonella enterica serovar Typhi strains isolated from Nepal in 2002 and 2003

A total of 171 Salmonella enterica serovar Typhi strains isolated from Nepal, mostly from patients with typhoid fever in 2002-2003, were tested for antimicrobial susceptibility by disk diffusion assay. Selected S. enterica serovar Typhi isolates were tested for MICs by E-test for ceftriaxone, ciprofloxacin and ofloxacin. Mutations of DNA gyrase gyrA and gyrB and topoisomerase IV parC and parE were identified by sequencing of PCR amplicons. By disk diffusion assay, 75/171 S. enterica serovar Typhi isolates were resistant to nalidixic acid, ampicillin, choramphenicol, streptomycin, tetracycline, sulfisoxazole, and trimethroprim/sulfamethoxazoles. Multiple drug resistance to the 7 antimicrobials was most predominant among S. enterica serovar Typhi isolates in this study. Resistance to nalidixic acid was detected in 76/111 and 56/60 of total isolates collected in 2002 and 2003, respectively. Nalidixic acid-resistant isolates in 2002 and 2003 showed MIC range for ciprofloxacin of 0.125-0.250 mg/l. Nalidixic acid-resistant isolates contained point mutations in gyrA and parC but not gyrB and parE. The gyrA mutation of nalidixic acid-resistant isolates obtained in 2002 and 2003 had amino acid substitution at position 83 of Serine-->Tyrosine and Serine-->Phenylalanine, respectively. Two different mutations of gyrA were detected among nalidixic acid-resistant isolates. Thus it is necessary to monitor mutation in DNA topoisomerase associated with increases in quinolones resistance.     

Quinolone-resistant Escherichia coli

Quinolones (nalidixic acid - NAL, norfloxacin - NOR, ciprofloxacin - CIP and gatifloxacin - GAT) were tested against Escherichia coli isolated from urine (385 patient samples) by disk diffusion (DD) and agar dilution (AD) methods. Fifty-three samples (13.8%) were classified as resistant to at least one of the quinolones tested. CIP and NOR susceptibilities were the same (91.4%) and they were similar to GAT (92.7%). Susceptibility to NAL, detected by the disk diffusion method, was used to predict susceptibility to NOR, CIP and GAT by the agar dilution method. The sensitivity and specificity of NAL were 100% and 95%, respectively. Twelve samples were analyzed for mutations in the quinolone resistance-determining region (QRDR) of the gyrA and parC genes. Sequencing of these genes failed to find any mutations in the quinolone-sensitive isolates. However, three mutations were observed in the isolates resistant to all the quinolones tested - two in gyrA and one in parC. A single mutation in gyrA was found in the strains that were resistant to nalidixic acid but fluoroquinolone-sensitive. These findings support the suggestion that NAL could be used as a marker for susceptibility to fluoroquinolones in routine microbiology laboratories. The overall resistance rate to quinolones in the present study was 13.8%, which is higher than that observed in other studies carried out in developed countries. Our findings serve as a warning that resistance to this group of antimicrobial agents is increasing.     

Antibiotic susceptibility pattern and the indicator of decreased ciprofloxacin susceptibility of Salmonella enterica serovar Typhi isolated from Dhulikhel Hospital, Nepal

Monitoring the antibiotic susceptibility pattern of Salmonella enterica serovar Typhi (S. Typhi) is important for efficiently managing cases of typhoid fever. In this study, the antimicrobial susceptibility patterns of 114 S. Typhi isolates, which were collected from a university hospital in Nepal during July 2009-December 2010, were investigated by disc diffusion assays. All of the S. Typhi isolates were sensitive to amoxycillin-clavulanic acid. More than 95% of the isolates were sensitive to chloramphenicol, ceftazidime, ceftriaxone, and cotrimoxazole. In addition, 1.7% of the studied isolates showed multiple drug resistance patterns. Of the 40 S. Typhi isolates, 32 strains (80%) showed nalidixic acid (NA) resistance with decreased susceptibility to ciprofloxacin (CIP). Importantly, we found the simultaneous presence of NA resistance and decreased susceptibility to CIP, suggesting that the resistance to NA is a reliable indicator of decreased CIP susceptibility (sensitivity, 97.5%; specificity, 100.0%). Furthermore, the sequencing of NA-resistant S. Typhi isolates showed a predominant amino acid alteration in the quinolone resistance-determining region (QRDR) of gyrA gene at position 83 from Ser→Phe. Two isolates with resistance to both CIP and NA had a double-mutation (Ser83→Phe and Asp87→Asn) in the QRDR of the gyrA gene, of which one had an additional amino acid mutation (Ser80→Ilu) in the QRDR of the parC gene.     

耐多药结核分枝杆菌对喹诺酮类药物的耐药性与gyr基因突变的初步研究

目的分析耐多药结核分枝杆菌（Multiple drug-resistant tuberculosis,MDR-TB）临床分离株的gyr基因突变特点,探讨MDR-TB对喹诺酮类药物耐药产生与gyr基因突变的关系。方法采用比例法对耐多药结核临床分离菌株进行氧氟沙星的药物敏感性检测,应用DNA直接测序法检测MDR-TB的gyr基因突变情况。结果 125株MDR-TB临床分离株中,50株对喹诺酮类耐药,耐药率为40%。50株耐药菌株中,40株gyr基因发生突变：其中39株gyrA基因突变,突变率为78%,突变位点包括90,91和94位氨基酸;5株gyrB基因突变,其中4株合并gyrA基因突变,gyrB基因突变位点为500,506,534和539位氨基酸。结论 MDR-TB中的喹诺酮类药物耐药态势比较严峻,其对喹诺酮类药物耐药机制主要与gyrA基因突变有关。     

Serotypes and antimicrobial resistance of Campylobacter jejuni isolated from humans and animals in Thailand

We investigated the serotypes, distributions, and antimicrobial resistance of Campylobacter jejuni isolates from humans and animals as a source of infection in poultry between 2002 and 2003. A total of 50 C. jejuni isolates from humans and 29 C. jejuni isolates from poultry were studied for serotype using the Penner serotyping scheme and the drug susceptibilities of the isolates which were determined for 7 antimicrobial drugs using the disk diffusion method. Serotype B (10%), serotype E (8%) and serotype R (8%) were found in humans isolates, while serotype A (27%) was most freguently isolated from poultry, followed by serotype K (21%) and serotype C (13%). Resistance in human isolates to cephalothin was high (100%). Resistance to trimethoprim/sulfamethoxazole, ciprofloxacin, and nalidixic acid were observed in 90, 82 and 78% of isolates, respectively. Most of the isolates (88%) were susceptible to erythromycin. High levels of resistance to drugs (ciprofloxacin and nalidixic acid) were observed in the isolates from poultry. These results indicate the importance of poultry as a reservoir of C. jejuni infection in Thailand is limited. In addition, a high proportion of the isolates were resistant to antimicrobial drugs, particularly the quinolone group.     

Mutations in the gyrA and gyrB genes of fluoroquinolone-resistant Mycobacterium tuberculosis from TB patients in Thailand

Among fluoroquinolone-resistant Mycobacterium tuberculosis (FQr-MTB) isolates, mutation at positions 90, 91, and 94 in gyrA gene and at positions 495, 516, and 533 in gyrB gene have been frequently reported. In this study, 35 isolates of FQr-MTB were collected from Siriraj Hospital and Chest Disease Institute. The quinolone-resistance-determining regions (QRDR) of gyrA and gyrB genes in all 35 FQr-MTB isolates and from the H37Ra MTB strain were amplified using polymerase chain reaction (PCR). DNA-sequencing and single-strand conformation polymorphism (SSCP) were further utilized for characterization of the mutations in the QRDR of gyrA and gyrB genes and mutation screening, respectively. From DNA-sequencing, 21 of 35 (60%) exhibited single-point mutations in different positions, at Ala90Val, Ser91Pro, and Asp94(Gly/Ala/His/Asn); and one novel mutation position at Gly88Cys in the gyrA gene and Asp495Asn in the gyrB gene. These positions were previously frequently reported to be responsible for FQr-MTB. The other 14 FQr-MTB isolates (40%) had no mutation. This study is the first report of mutation occurring only in the QRDR of the gyrB gene, without prior mutation in the gyrA QRDR among FQr-MTB isolates. By SSCP analysis for screening of the mutant FQr-MTB, the SSCP patterns of mutated FQr-MTB isolates were clearly differentiated from the SSCP patterns of FQs-MTB.     

Molecular epidemiology of fluoroquinolone-resistant Streptococcus pneumoniae in Japan

We identified fluoroquinolone-resistant Streptococcus pneumoniae strains among 670 clinical isolates isolated from 1999 to 2003 in Hokkaido prefecture, Japan. All eleven stains were resistant to ciprofloxacin and levofloxacin. Furthermore, ten strains were also resistant to fluoroquinolones that are more effective with gram-positive bacteria, namely tosufloxacin, sparfloxacin, and gatifloxacin. Nucleotide sequence analysis of the quinolone-resistance determining region (QRDR) of the quinolone target genes coding for topoisomerase i.v. subunits (parC and parE) and DNA gyrase subunits (gyrA and gyrB). Eight stains, which showed higher resistance, had resistance mutations in two genes (gyrA and parC, or gyrA and parE), and other three strains had one resistance mutation in parC. The mutation patterns were varied between the strains. Data from random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) indicated that eleven strains were identified as ten independent clones. Lines of evidence indicated that genetic mutations leading to fluoroquinolone resistance occur sporadically rather through the spreading of a particular resistant strain. Notably, the fluoroquinolone-resistant strains were only isolated from adults, particularly from patients more than 60 years of age (9/60 strains; 15.0%). Resistant strains were not found in 574 strains isolates from patients under 20 years of age. This may be due to the fact that fluoroquionolones other than norfloxacin are not applicable to children in Japan.     

A possible mechanism of macrolide resistance among multiple resistant Campylobacter jejuni and Campylobacter coli isolated from Thai children during 1991-2000

A total of 495 Campylobacterjejuni and 122 C. coli isolated from Thai children were screened for macrolide (erythromycin and azithromycin) resistance by disk diffusion assay. Minimum inhibitory concentrations for erythromycin, azithromycin, nalidixic acid, ciprofloxacin, tetracycline, streptomycin, gentamicin and chloramphenicol were further determined for these macrolide-resistant Campylobacter isolates. Presence of known point mutations resulting in reduced susceptibility to macrolides was investigated by PCR and DNA sequencing. Seventeen percent (23/122) of C. coli and 2.4% (12/495) of C. jejuni isolates were resistant to macrolides. By sequencing domain V of the 23S ribosomal DNA from all 35 macrolide-resistant isolates, a known point mutation of 23S rRNA associated with reduced susceptibility to macrolides was detected in all isolates except one. Among the macrolide-resistant isolates, all were multiply resistant to nalidixic acid and ciprofloxacin, of which the latter is the preferred antimicrobial used for diarrheal treatment in Thailand. Furthermore, most macrolide-resistant isolates were also resistant to tetracycline and streptomycin. The spread of macrolide and quinolone resistant Campylobacter should be monitored closely in Thailand and elsewhere as these antimicrobials are preferred drugs for treatment of diarrhea.