摘要
目的评价脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)技术用于肺炎克雷伯菌(Klebsiella pneumoniae)的分型能力。方法选用220株肺炎克雷伯菌,对PFGE方法的分型力、可重复性、分辨力、流行病学一致性及其与多位点序列分型(MLST)的分型一致性进行评价。挑选全部220株菌株进行实验,评价PFGE的分型力;挑选18株菌株重复2次实验评价PFGE的可重复性;分别挑选3组无流行病关联的碳青霉烯耐药临床株、腹泻患者肠道分离株、食品分离株,使用Simpson差异指数(D)评价PFGE的分辨力;挑选同一次暴发期间分离的34株菌株,评价PFGE的流行病学一致性及其与MLST对暴发菌株分型结果的一致性;挑选64株不同来源的菌株,评价PFGE和MLST分型对流行病学不相关菌株的分型一致性。用Bio Numerics软件对所有实验菌株的PFGE图谱进行分析。结果 220株肺炎克雷伯菌中有216株通过PFGE能够获得有效的图谱,其分型力为98.18%(216/220);18株菌株重复2次实验的图谱完全一致;PFGE对无流行病关联的50株碳青霉烯耐药临床株、34株腹泻患者肠道分离株、48株食品分离株分型的D值分别为0.9910、0.9964、1.0000;针对34株暴发期间分离菌株,PFGE能够很好地甄别出暴发菌株,高度相关菌株和不相关菌株,并且分型结果与菌株分离病区、分离时间具有一致性;针对暴发菌株,PFGE能够将相同MLST型别的菌株聚成一簇;针对流行病学不相关菌株,PFGE不能将相同MLST型别的菌株分成相同或相似的带型或者聚成一簇。结论 PFGE对肺炎克雷伯菌分型具有很好的分型力、可重复性和分辨力;针对暴发菌株,PFGE能够将相同MLST型别的菌株聚成一簇;针对流行病学不相关菌株,PFGE和MLST分型一致性差。
Objective To evaluate the performance of pulsed field gel electrophoresis（ PFGE） in the subtyping of Klebsiella pneumoniae. Methods PFGE was used to detect 220 K. pneumoniae strains to evaluate its subtyping capability and reproducibility; three groups of epidemiologic unlinked strains of carbapenemase-producing clinical strains,diarrhea pathogen strains and foodborne strains were used to evaluate PFGE discriminatory power,34 strains isolated during an outbreak were used to evaluate epidemiologic concordance and the subtyping consistency between PFGE and multilocus sequence typing（ M LST） and 64 strains from different sources were used to evaluate the subtyping consistency of epidemiologic unlinked strains between PFGE and M LST. Bio Numerics was used to develop the molecular typing database based on PFGE patterns. Results The PFGE asasay used in this study had high subtyping capability（ 98. 18%）,reproducibility（ 100%） and discriminatory power（ D = 0. 9910,0. 9964,1. 0000 for three groups of epidemiologic unlinked strains respectively）. For 34 strains isolated during an outbreak,PFGE identified the strains causing the outbreak and unlinked strains,and had good subtyping consistency with M LST. However,PFGE could not subtype and cluster the strains with same M LST types. Conclusion PFGE has good subtyping capability,reproducibility,discriminatory power,epidemiologic consistency in detecting K. pneumoniae strains; PFGE had good subtyping consistency with M LST for outbreak strains,but the consistency was poor between PFGE and M LST for epidemiologic unlinked strains.
出处
《疾病监测》
CAS
2015年第11期969-975,共7页
Disease Surveillance
基金
国家"十一五"科技重大专项(No.2008ZX10004-008)
国家"十二五"科技重大专项(No.2012ZX10004215)~~
关键词
肺炎克雷伯菌
脉冲场凝胶电泳
多位点序列分型
分子分型
Klebsiella pneumoniae
Pulsed field gel electrophoresis
Multilocus sequence typing
Molecular typing
