铜绿假单胞菌随机扩增多态性DNA指纹法基因分型研究

Genotyping of Pseudomonas aeruginosa by randomly amplified polymorphic DNA fingerprinting

建立了铜绿假单胞菌（ＰＡ）随机扩增多态性ＤＮＡ（ＲＡＰＤ）指纹图基因分型方法，并应用于流行病学相关或不相关的７５个ＰＡ菌株的基因分型。分型率为１００％。３２株新生儿暴发感染菌株，可分成４个血清型和５个ＲＡＰＤ谱型，其中２５株属于同一谱型。在８例患者的垂直追踪菌株中，除有１例其第１和第３个分离株的血清型和ＲＡＰＤ谱型均相同，但与第２个分离株的血清型和ＲＡＰＤ谱型不同外，其余７例前后菌株的谱型相同。１３株散发菌株可分成３个血清型和１０个ＲＡＰＤ谱型；１２株流行病学无关菌株的ＲＡＰＤ谱型均不相同。本研究结果表明，ＲＡＰＤ指纹图基因分型法具有分型率高、分辨力强、比较快速简便等优点，并且不需要已知核酸序列，是在分子水平上对微生物感染的病原学、发病机理及流行病学研究的较理想的方法。

Randomly amplified polymorphic DNA fingerprinting was established and used to type 75 epidemiological related or unrelated strains of Pseudomonas aeruginosa .Typeability was 100%.Thirty two epidemic strains from the newborns were divided into 4 serotypes and 5 RAPD profiles;among which 25 strains had a same profile.The first and the third isolate from patient C were the same both in serotype and RAPD profile,but different from the second one both in serotype and RAPD profile.The follow up strains from each of 7 other patients had a same profile.Thirteen sporadic isolates were divided into 3 serotypes and 10 RAPD profiles.Twelve epidemiological unrelated strains were distinguishable each other in RAPD profiles.The results showed that RAPD fingerprint genotyping is suitable for studies on the etiology,pathogenesis and epidemiology of microbe infection at molecular level with high typeability,powerful discrimination,rapidness and simplicity.