Zhang Wen-li, Liu Jun, Zhang Jian, Su Guang-long
Department of Clinical Laboratory, The Children's Hospital, Urumqi, Urumqi 830002, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2011 Mar;45(3):217-24.
This study aimed to investigate drug resistance and genotypes of the extended spectrum β-lactamase (ESBLs) and AmpC β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolated from Uygur and Han newborns in Urumqi.
Disk diffusion test (Kirby-Bauer) was used for detecting drug resistance of 299 strains to twenty two kinds of antibiotics. Resistance genes of the ESBLs and AmpC β-lactamase-producing strains were amplified by multiplex PCR and subtypes were confirmed by DNA sequence analysis. Total 148 strains were selected with random number table and sequenced, which included TEM-, SHV-, CTX-M-1-, or CTX-M-9-positive ESBLs-producing strains and DHA-, or CIT-positive AmpC β-lactamase-producing strains. Antibiotic resistant rates were analyzed by Whonet 5.4 and statistic analysis was performed by chi-square (χ(2)) test with PEMS 3.1.
The antibiotic resistant rates between Uygur and Han newborns significantly differ in ESBLs-producing Klebsiella pneumoniae to Sulfamethoxazole-Trimethoprim (80.0% (40/50) and 56.0% (28/50), χ(2) = 6.6176, P = 0.0101), in ESBLs-producing Escherichia coli to Sulbactam and Cefoperazone (54.2% (32/59) and 94.0% (47/50), χ(2) = 21.4512, P = 0.0000), and in AmpC β-lactamase-producing Klebsiella pneumoniae to Sulbactam and Cefoperazone (100.0% (20/20) and 72.2% (26/36), χ(2) = 6.7633, P = 0.0093) and to Amikacin (65.0% (13/20) and 25.0% (9/36), χ(2) = 8.6246, P = 0.0033). Although SHV gene of ESBLs-producing Escherichia coli was detected from Uygur newborns at only 3.4% (2/59) and not detectable from Han newborns, TEM, CTX-M-1, and CTX-M-9 group genes were all detected over 38.0% (19/50). Among the detected strains, the subtypes of TEM and CTX-M-1 were mainly TEM-1 and CTX-M-15, respectively; whereas the subtypes of SHV and CTX-M-9 included SHV-1, 2, 11, 12, 27, 61, 99 and CTX-M-9, 14, 24, 27, 65, respectively. The strains of Escherichia coli and Klebsiella pneumoniae carrying two or more kinds of ESBLs genotypes were 56.7% (42/74) - 90.0% (63/70). Two species carrying the AmpC gene in two kinds of newborns were only grouped in the subtypes of DHA-1 and CMY-44, and other subtypes were not detected at all. Moreover, TEM-positive ESBLs-producing Escherichia coli were detected from Uygur newborns at the higher rate than that from Han newborns (71.2% (42/59) and 50.0% (25/50), χ(2) = 5.1291, P = 0.0235), while there was no difference in other genotypes detected between two kinds of newborns (χ(2) < 3.7780, P > 0.05).
There were significant differences in antibiotic resistance and genotype distribution of Klebsiella pneumoniae and Escherichia coli between two nationality newborns, and these two bacteria detected in this study carried multi-resistance genes and showed high resistant to β-lactamase antibiotics.
本研究旨在调查从乌鲁木齐维吾尔族和汉族新生儿中分离出的产超广谱β-内酰胺酶(ESBLs)和AmpCβ-内酰胺酶的大肠埃希菌及肺炎克雷伯菌的耐药性和基因型。
采用纸片扩散法(Kirby-Bauer)检测299株菌对22种抗生素的耐药性。通过多重聚合酶链反应(PCR)扩增产ESBLs和AmpCβ-内酰胺酶菌株的耐药基因,并用DNA序列分析确定亚型。用随机数字表法共选取148株菌进行测序,其中包括产TEM-、SHV-、CTX-M-1-或CTX-M-9的ESBLs菌株及产DHA-或CIT的AmpCβ-内酰胺酶菌株。用Whonet 5.4分析抗生素耐药率,并用PEMS 3.1软件进行χ²检验的统计学分析。
维吾尔族和汉族新生儿中产ESBLs的肺炎克雷伯菌对复方磺胺甲恶唑的耐药率有显著差异(80.0%(40/50)和56.0%(28/50),χ² = 6.6176,P = 0.0101);产ESBLs的大肠埃希菌对舒巴坦和头孢哌酮的耐药率有显著差异(54.2%(32/59)和94.0%(47/50),χ² = 21.4512,P = 0.0000);产AmpCβ-内酰胺酶的肺炎克雷伯菌对舒巴坦和头孢哌酮及阿米卡星的耐药率有显著差异(100.0%(20/20)和72.2%(26/36),χ² = 6.7633,P = 0.0093)及(65.0%(13/20)和25.0%(9/36),χ² = 8.6246,P = 0.0033)。虽然维吾尔族新生儿中产ESBLs的大肠埃希菌SHV基因检出率仅为3.4%(2/59),汉族新生儿未检出,但TEM、CTX-M-1和CTX-M-9组基因在两组中的检出率均超过38.0%(19/50)。在检出的菌株中,TEM和CTX-M-1亚型分别主要为TEM-1和CTX-M-15;而SHV和CTX-M-9亚型分别包括SHV-1、2、11、12、27、61、99和CTX-M-9、14、24、27、65。携带两种或以上ESBLs基因型的大肠埃希菌和肺炎克雷伯菌菌株比例为56.7%(42/74)-90.0%(63/70)。两种新生儿中携带AmpC基因的两种菌仅分型为DHA-1和CMY-44亚型,未检出其他亚型。此外产TEM的ESBLs大肠埃希菌在维吾尔族新生儿中的检出率高于汉族新生儿(71.2%(42/59)和50.0%(25/50),χ² = 5.1291,P = 0.0235),而其他基因型在两组中的检出差异无统计学意义(χ² < 3.7780,P > 0.05)。
两个民族新生儿肺炎克雷伯菌和大肠埃希菌的耐药性及基因型分布存在显著差异,本研究检出的这两种菌携带多种耐药基因,对β-内酰胺类抗生素呈现高度耐药。