Hansen D S, Skov R, Benedí J V, Sperling V, Kolmos H J
The International Escherichia and Klebsiella Reference Center (WHO), Statens Serum Institut, Copenhagen, Denmark.
Clin Microbiol Infect. 2002 Jul;8(7):397-404. doi: 10.1046/j.1469-0691.2002.00411.x.
To compare pulsed-field gel electrophoresis (PFGE) typing and O:K-serotyping of Klebsiella in two different epidemiological settings.
One hundred and four bacteremia isolates without known epidemiological relation and 47 isolates from an outbreak in a neonatal intensive care unit (NICU) were K-typed by countercurrent immunoelectrophoresis (CCIE), O-typed by an inhibition enzyme-linked immunosorbent assay method, and typed by pulsed-field gel electrophoresis (PFGE) using the restriction enzyme XbaI.
Typing data for the 104 bacteremia isolates were compared with regard to typability, number of types, maximum number of isolates per type, and the Discriminative Index (DI). O-typing combined with K-typing (DI 0.98) as O:K-serotyping (DI 0.99) gave a very discriminative typing system, whereas O-typing alone was not very discriminative (DI 0.76). PFGE (DI 1.00) was a more discriminative typing method than O:K-serotyping, as it could subdivide 13/22 O:K-serotypes into smaller groups. Isolates with the same PFGE-type had the same O:K-serotype, indicating that isolates with different O- and/or K-types could be expected to be of different PFGE-types. Typing of the 47 isolates from the outbreak in the NICU showed that 38 isolates belonged to a single clone, and that during an epidemic limited in time and space, differences in the electrophoretic patterns of up to five bands between a parental pattern type and a subtype may be found in the PFGE profiles.
Both O:K-serotyping and PFGE typing are highly discriminative typing methods. PFGE is the most discriminative method and is excellent for typing outbreaks with few isolates. If large numbers of isolates are to be typed, a more convenient strategy might be first to K- or O:K-serotype isolates followed by PFGE typing of possible identical isolates. Since K- or O:K-serotyping is a definitive typing method, while PFGE typing is a comparative one, PFGE cannot, for the time being, replace O:K-serotyping for surveillance purposes.
比较两种不同流行病学背景下肺炎克雷伯菌的脉冲场凝胶电泳(PFGE)分型和O:K血清分型。
采用对流免疫电泳(CCIE)对104株无已知流行病学关联的菌血症分离株及47株来自新生儿重症监护病房(NICU)暴发的分离株进行K分型,采用抑制酶联免疫吸附测定法进行O分型,并用限制性内切酶XbaI通过脉冲场凝胶电泳(PFGE)进行分型。
比较了104株菌血症分离株的分型数据,包括可分型性、型别数量、每种型别的最大分离株数量以及鉴别指数(DI)。O分型与K分型相结合(DI 0.98),即O:K血清分型(DI 0.99),形成了一个鉴别力很强的分型系统,而单独的O分型鉴别力不强(DI 0.76)。PFGE(DI 1.00)是一种比分型O:K血清分型更具鉴别力的分型方法,因为它可以将13/22种O:K血清型细分为更小的组。具有相同PFGE型别的分离株具有相同的O:K血清型,这表明具有不同O和/或K型别的分离株可能具有不同的PFGE型别。对NICU暴发的47株分离株进行分型显示,38株属于单一克隆,并且在时间和空间有限的疫情期间,在PFGE图谱中可能会发现亲代型别和亚型之间的电泳图谱差异多达五条带。
O:K血清分型和PFGE分型都是高鉴别力的分型方法。PFGE是最具鉴别力的方法,对于分离株较少的暴发分型非常适用。如果要对大量分离株进行分型,一种更便捷的策略可能是首先对分离株进行K或O:K血清分型,然后对可能相同的分离株进行PFGE分型。由于K或O:K血清分型是一种确定性分型方法,而PFGE分型是一种比较性分型方法,目前PFGE不能替代O:K血清分型用于监测目的。