不同来源奇异变形杆菌对小鼠致病力及其毒力基因携带情况的比较
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摘要
目的比较通辽地区不同来源奇异变形杆菌对小鼠致病力及其毒力基因携带情况。方法取4株(SPM、GPM、BPM-1和BPM-2)不同来源的奇异变形杆菌,进行生化鉴定、药物敏感性测试及动物致病性试验,对4株菌株的8种毒力基因(ureC、zapA、mrpA、ucaA、rsbA、pmfA、atfA、atfC)进行PCR检测,对其16S r RNA基因序列进行同源性和系统进化树分析。结果 4株菌株均被鉴定为奇异变形杆菌;对4株病原菌均高度敏感的药物有丁胺卡那、美罗培南、头孢他啶、头孢噻肟、头孢吡肟、他巴克坦和氨曲南7种,均不敏感的药物有庆大霉素、亚胺培南、头孢唑啉、氨苄西林、克拉维酸、黏菌素、磺胺甲恶唑、氯霉素、环丙沙星和四环素10种;4株菌株的半数致死量(LD50)按BPM-2、BPM-1、SPM、GPM依次降低,鹅源的毒力最高;奇异变形杆菌具有不同的毒力基因谱,8种毒力基因在菌株SPM、GPM、BPM-2中均被检出,菌株BPM-2未检出ucaA基因。BPM-2和BPM-1处于同一分支,但分处于不同的末支;SPM和GPM分处于不同的小分支。结论不同来源的奇异变形杆菌对小鼠的致病力存在较小的差异,且对小鼠有致病力的基因表型与该毒力基因的分布可能存在一定的相关性。
Objective To compare the pathogenicity in mice and carriage of virulence genes of Proteus mirabilis isolates from various origins in Tongliao Region,Inner Mongolia university for Nationalities,China. Methods Four strains of Proteus mirabilis from various origins,SPM,GPM,BPM-1 and BPM-2,were subjected to biochemical identification,drug sensitivity test and pathogenicity test in animals,of which the eight virulence genes(ureC,zapA,mrpA,ucaA,rsbA,pmfA,atf A and atfC) were detected by PCR. The 16 S r RNA gene sequences of four strains were analyzed for homology,based on which a phylogenetic tree was plotted. Results All the four strains were identified as P. mirabilis,to which the highly sensitive drugs included amikacin,meropenem,ceftazadine,cefotaxime,cefepime,taxoeillin and aztreonam. However,gentamicin,imipenem,cefazolin,ampicillin,clavulanic acid,colysin,sulfamethoxazole,chloramphenicol,ciprofloxacin and tetracycline were insensitive to any of the four strains. In the turn of median lethal dose,the four strains were BPM-2,BPM-1,SPM and GPM,while the virulence of GPM was the highest. The P. mirabilis strains showed different virulence gene profiles. All the eight virulence genes were detected in SPM,GPM and BPM-2,while ucaA gene was undetected in BPM-2. BPM-2 and BPM-1 belonged to the difference branches of the same clade.However,SPM and GPM belonged to difference clades. Conclusion P. mirabilis strains from different origins showed minor difference in pathogenicity to mice. The phenotype of pathogenic virulence gene might be correlated to the distribution of the gene.
Objective To compare the pathogenicity in mice and carriage of virulence genes of Proteus mirabilis isolates from various origins in Tongliao Region,Inner Mongolia university for Nationalities,China. Methods Four strains of Proteus mirabilis from various origins,SPM,GPM,BPM-1 and BPM-2,were subjected to biochemical identification,drug sensitivity test and pathogenicity test in animals,of which the eight virulence genes(ureC,zapA,mrpA,ucaA,rsbA,pmfA,atf A and atfC) were detected by PCR. The 16 S r RNA gene sequences of four strains were analyzed for homology,based on which a phylogenetic tree was plotted. Results All the four strains were identified as P. mirabilis,to which the highly sensitive drugs included amikacin,meropenem,ceftazadine,cefotaxime,cefepime,taxoeillin and aztreonam. However,gentamicin,imipenem,cefazolin,ampicillin,clavulanic acid,colysin,sulfamethoxazole,chloramphenicol,ciprofloxacin and tetracycline were insensitive to any of the four strains. In the turn of median lethal dose,the four strains were BPM-2,BPM-1,SPM and GPM,while the virulence of GPM was the highest. The P. mirabilis strains showed different virulence gene profiles. All the eight virulence genes were detected in SPM,GPM and BPM-2,while ucaA gene was undetected in BPM-2. BPM-2 and BPM-1 belonged to the difference branches of the same clade.However,SPM and GPM belonged to difference clades. Conclusion P. mirabilis strains from different origins showed minor difference in pathogenicity to mice. The phenotype of pathogenic virulence gene might be correlated to the distribution of the gene.
引文
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[20]姚金元,杨一鸣,顾蓓青,等.135株奇异变形杆菌的标本分布及耐药性分析[J].中国消毒学杂志,2016,33(3):289-290.
[21]FENG F Y,YANG X Y,HONG N,et al.Study on distribution of drug resistance gene and integron and analysis of genetic relationship of 20 isolates of Proteus mirabilis[J].Int J Lab Med,2015,36(17):2461-2463.(in Chinese)冯福英,杨湘越,洪宇,等.20株奇异变形杆菌耐药基因和整合子分布及亲缘关系分析[J].国际检验医学杂志,2015,36(17):2461-2463.
[22]LUO S,LIU W E,YAN Q,et al.Clinical distribution and antimicrobial resistance of 172 Proteus mirabilis isolates and 68Proteus vulgaris isolates[J].Chin J Infect Control,2014,13(12):710-713.(in Chinese)罗珊,刘文恩,晏群,等.172株奇异变形杆菌和68株普通变形杆菌临床分布及其耐药性[J].中国感染控制杂志,2014,13(12):710-713.
[23]CHENG Q,PANG R L,WANG R C,et al.Comparative study on pathogenicity of Salmonella isolates from different sources of laboratory mice and the detection of their virulence genes[J].Chin J Zoon,2013,29(5):460-465.(in Chinese)程琼,庞瑞亮,王若晨,等.不同源沙门氏菌对小鼠致病力的比较与毒力基因检测[J].中国人兽共患病学报,2013,29(5):460-465.
[2]刘燕云,柴家前.奇异变形杆菌研究进展[J].中国兽医学报,2017,37(1):196-200.
[3]BISGAARD M,DAM A.Salpingitis in poultry.II.Prevalence,bacteriolo gy and possible pathogenesis in egg-laying chickens[J].Nord Vet Med,1981,33(2):81-89.
[4]江益民,龙塔,侯玉泽,等.鸡奇异变形杆菌病病原的分离与鉴定[J].河南农业科学,1990,19(7):31-33.
[5]王岩,尹珺伊,侯美如,等.鹅源奇异变形杆菌的分离鉴定及药敏试验[J].中国家禽,2017,39(23):56-59.
[6]马婷婷,韦显凯,闭璟珊,等.猪源奇异变形杆菌的分离鉴定及其毒力的测定[J].中国兽医科学,2017,47(10):1234-1239.
[7]LI Y X,JIAO F C,PAN Z M,et al.Identification and genetic evolution analysis of Proteus mirabilis isolated from peacocks[J].Anim Husb Vet Med,2016,48(5):79-81.(in Chinese)李迎晓,焦凤超,潘志明,等.孔雀奇异变形杆菌的分离鉴定及遗传进化分析[J].畜牧与兽医,2016,48(5):79-81.
[8]WANG L P,MENG Y,LV J Y,et al.Isolation and identification of mink Proteus mirabilis[J].Heilongjiang Anim Sci Vet Med,2016,59(5):164-166,288.(in Chinese)王丽萍,孟洋,吕九云,等.水貂奇异变形杆菌的分离鉴定[J].黑龙江畜牧兽医,2016,59(5):164-166,288.
[9]LIN Y G,YE J,SHI T T,et al.Isolation and identification of pathogen Proteus mirabilis from diseased Chinese soft-shelled turtle Pelodiscus sinensis[J].Fish Sci,2014,33(12):800-803.(in Chinese)林亚歌,叶键,石婷婷,等.中华鳖源奇异变形杆菌的分离鉴定与致病性研究[J].水产科学,2014,33(12):800-803.
[10]FAN K W,YU J,YANG S S,et al.Isolation,identification and phylogenetic analysis of Proteus mirabilis from South China tiger[J].Vet Sci in China,2018,48(1):69-75.(in Chinese)范克伟,余佳,杨守深,等.华南虎源奇异变形杆菌的分离鉴定及其系统发育分析[J].中国兽医科学,2018,48(1):69-75.
[11]XIONG H M,WEI B Q,WEI R J,et al.Calculation of median lethal dose(LD50)for Yersinia pesos by SPSS package[J].Chin J Zoon,2013,29(11):1127-1130.(in Chinese)熊浩明,魏柏青,魏荣杰,等.用SPSS软件计算鼠疫菌半数致死量(LD50)[J].中国人兽共患病学报,2013,29(11):1127-1130.
[12]ZHU F Z,CHEN L Y,CHEN H C.Identification of pathogenic microorganismby sequencing 16S rRNA gene[J].J Cent South Univ(Med Sci),2013,38(10):1035-1041.(in Chinese)朱飞舟,陈利玉,陈汉春.16S rRNA基因序列分析法鉴定病原细菌[J].中南大学学报(医学版),2013,38(10):1035-1041.
[13]YIN Y Q,MA D H,LI Y S,et al.Isolation,identification of goose Proteus mirabilis and it's virulence genes detection[J].Microbiol China,2018,45(10):2152-2163.(in Chinese)尹有勤,马德慧,李岩松,等.鹅源奇异变形杆菌的分离鉴定及毒力基因检测[J].微生物学通报,2018,45(10):2152-2163.
[14]YANG Q L,G J,LUO L P,et al.Virulence tests of canine Proteus mirabilis[J].Chin J Vet Med,2013,49(8):60-63.(in Chinese)杨巧玲,高健,罗立平,等.犬源性奇异变形杆菌的毒力试验[J].中国兽医杂志,2013,49(8):60-63.
[15]LU Z X,SUN K,GUO W N,et al.Isolation and identification of Proteus and detection of virulence-and resistant-associated genes[J].Anim Husb Vet Med,2017,49(12):97-101.(in Chinese)路振香,孙坤,郭伟娜,等.变形杆菌的分离鉴定及其毒力与耐药相关基因检测[J].畜牧与兽医,2017,49(12):97-101.
[16]NAKAMA R,SHINGAKI A,MIYAZATO H,et al.Current status of extended spectrumβ-lactamase-producing Escherichia coli,Klebsiella pneumoniae and Proteus mirabilis in Okinawa prefecture,Japan[J].J Infect Chemother,2016,22(5):281-286.
[17]LI W J,LIU Y H,ZHOU C L,et al.Distribution and drug resistance of Proteus mirabilis[J].Chin J Nosocomiol,2017,27(12):2660-2663.(in Chinese)李文静,刘晔华,周春雷,等.奇异变形菌分布与耐药性分析[J].中华医院感染学杂志,2017,27(12):2660-2663.
[18]GUO Y M,PAN Z,QIN L Y,et al.Detection of virulence genes and analysis of drug sensitivity in Proteus mirabilis[J].Chin J Health Lab Tec,2016,26(18):2671-2673.(in Chinese)郭玉梅,潘琢,秦丽云,等.奇异变形杆菌毒力基因检测和药物敏感性分析[J].中国卫生检验杂志,2016,26(18):2671-2673.
[19]ZHANG L F,LI J,LU J X,et al.Progress in research of antibiotic resistance of Proteus mirabilis[J].Dis Surveil,2016,31(5):427-432.(in Chinese)张利锋,李娟,卢金星.奇异变形杆菌耐药性研究进展[J].疾病监测,2016,31(5):427-432.
[20]姚金元,杨一鸣,顾蓓青,等.135株奇异变形杆菌的标本分布及耐药性分析[J].中国消毒学杂志,2016,33(3):289-290.
[21]FENG F Y,YANG X Y,HONG N,et al.Study on distribution of drug resistance gene and integron and analysis of genetic relationship of 20 isolates of Proteus mirabilis[J].Int J Lab Med,2015,36(17):2461-2463.(in Chinese)冯福英,杨湘越,洪宇,等.20株奇异变形杆菌耐药基因和整合子分布及亲缘关系分析[J].国际检验医学杂志,2015,36(17):2461-2463.
[22]LUO S,LIU W E,YAN Q,et al.Clinical distribution and antimicrobial resistance of 172 Proteus mirabilis isolates and 68Proteus vulgaris isolates[J].Chin J Infect Control,2014,13(12):710-713.(in Chinese)罗珊,刘文恩,晏群,等.172株奇异变形杆菌和68株普通变形杆菌临床分布及其耐药性[J].中国感染控制杂志,2014,13(12):710-713.
[23]CHENG Q,PANG R L,WANG R C,et al.Comparative study on pathogenicity of Salmonella isolates from different sources of laboratory mice and the detection of their virulence genes[J].Chin J Zoon,2013,29(5):460-465.(in Chinese)程琼,庞瑞亮,王若晨,等.不同源沙门氏菌对小鼠致病力的比较与毒力基因检测[J].中国人兽共患病学报,2013,29(5):460-465.