广东省宠物源肺炎克雷伯菌耐药、毒力基因的检测与药物敏感性测定

Resistance and virulence genes and antimicrobial susceptibility of pet-derived Klebsiella pneumoniae from Guangdong Province

摘要:

目的 探究广东省部分地区宠物源肺炎克雷伯菌的耐药和毒力情况。方法 采集犬猫粪便拭子，通过分离培养、PCR扩增16S rRNA和khe基因进行菌株鉴定。采用琼脂扩散法测定肺炎克雷伯菌分离株对17种抗菌药物的敏感性；通过PCR检测β-内酰胺类(bla_SHV、bla_CTX、bla_TEM)、碳青霉烯类(bla_KPC、bla_NDM)、氨基糖苷类(rmtB)、喹诺酮类(qnrS、oqxAB)、磺胺类(Sul1、Sul2)、酰胺醇类(floR)、四环素类[tet(A)]、磷霉素(fosA3)耐药基因以及部分毒力基因(rmpA、maga、fimH、mrkD、uge、WabG、kfu、Aerobactin、ureA)。结果 从采集的428份粪便样品中分离得到126株肺炎克雷伯菌，分离率为29.4%。琼脂扩散法结果显示，126株分离菌对阿莫西林(75.40%)、氨苄西林(73.81%)、复方新诺明(61.90%)耐药率高，对头孢他啶、阿米卡星、安普霉素和恩诺沙星较为敏感，对替加环素、黏菌素、美罗培南敏感。耐药基因检测结果显示，磺胺类耐药基因oqxAB检出率最高，为86.51%；其次为β-内酰胺酶耐药基因bla_SHV (73.81%)、四环素类耐药基因tet(A) (52.68%)；其他耐药基因均有不同程度检出(0.79%-46.03%)，碳青霉烯类耐药基因bla_KPC、黏菌素耐药基因mcr-1和氨基糖苷类耐药基因rmtB未检出。毒力基因检测结果显示，尿素酶相关基因ureA检出率为100.00%，脂多糖相关基因uge检出率为95.54%，菌毛相关基因?mH的检出率为91.07%，其他毒力基因均有不同程度检出(2.70%-8.90%)，荚膜相关基因magA与菌毛相关基因mrkD未检出。结论 广东省部分地区宠物源肺炎克雷伯菌耐药状况严重，但致病性较弱，应加强对其耐药性与致病力的监测。此外，临床上应严格管理和合理使用抗生素，避免多重耐药肺炎克雷伯菌的产生与传播。

Abstract:

Objective To investigate the antibiotic resistance and virulence of pet-derived Klebsiella pneumoniae in some areas of Guangdong Province.Methods Fecal swabs were collected from dogs and cats for strain isolation, and 16S rRNA and khe genes were amplified by PCR to identify the strains. The sensitivity of K. pneumoniae isolates to 17 antibiotics was determined by the agar diffusion method. PCR was employed to detect resistance genes to β-lactams (bla_SHV, bla_CTX, and bla_TEM), carbapenems (bla_KPC and bla_NDM), aminoglycosides (rmtB), quinolones (qnrS and oqxAB), sulfonamides (Sul1 and Sul2), amphenicols (floR), tetracyclines (tet(A)), and fosfomycin (fosA3) and some virulence genes (rmpA, maga, fimH, mrkD, uge, WabG, kfu, Aerobactin, and ureA).Results A total of 126 strains of K. pneumoniae were isolated from 428 fecal samples, with an isolation rate of 29.4%. The 126 isolates had high resistance rates to amoxicillin (75.40%), ampicillin (73.81%), and cotrimoxazole (61.90%). They were moderately sensitive to ceftazidime, amikacin, apramycin, and enrofloxacin, and they were sensitive to tigecycline, colistin, and meropenem. The detection rate of the resistance gene oqxAB was the highest, which was 86.51%, followed by those of the β-lactam resistance gene bla_SHV (73.81%) and the tetracycline resistance gene tet(A) (52.68%). Other resistance genes were detected to varying degrees (0.79%-46.03%) and the carbapenem resistance gene bla_KPC, colistin resistance gene mcr-1, and aminoglycoside resistance gene rmtB were not detected. Among the virulence genes, the urease gene urea, lipopolysaccharide-related gene uge, and fimbria-related gene fimH showed the detection rates of 100.00%, 95.54%, and 91.07%, respectively. Other virulence genes were detected to varying degrees (2.70%-8.90%), while the capsule-related gene magA and the fimbria-related gene mrkD were not detected.Conclusion In some areas of Guangdong Province, the antibiotic resistance of pet-derived K. pneumoniae is serious, but its pathogenicity is relatively weak. Monitoring of its drug resistance and pathogenicity should be strengthened. In addition, antibiotics should be strictly managed and rationally used in the clinical practice, so as to avoid the generation and dissemination of multidrug resistant strains of K. pneumoniae.