角膜结膜及泪器病             1559                                   AB0358

　　Multiocus Sequencing Typing of Pseudomonas aeruginosa from bacterial keratitis

　　WangDan, LiangQingfeng,WangZhiqun ,LiRan, SunXuguang.

　　Beijing Institute of Ophthalmology, Beijing Tongren Ophthalmic Center,17 Hou Gou Lane,,Chong Nei Street Beijing  100005, China

　　Objective : To study on the genetic background of five  isolates of Pseudomonas aeruginosa from bacterial keratitis.

　　Methods: P. aeruginosa strains was from five patients(five eyes),all female. The range of age was from ten to seventy-one. The strains were cultured on BHI agar at 37 C under aerobic conditions.

　　Chromosomal DNA was extracted from these purified strains with a kit. PCR primers were designed specially for seven house-keeping loci: acsA, aroE, guaA,mulT,nuoD, ppsA, trpE. Amplification and sequencing of the loci, offer the sequencs to the website(http://www.pubmlst.org) The site assigned alleles at multiple house-keeping loci by sequencing. An arbitrary number was given to each distinct allele within a locus. Each isolate was therefore given seven numbers that represented its sequence type (ST).

　　Result:PA564 (43-30-31-26-4-24-32) belonged to ST127; PA583(17-24-37-5-4-7-7)belonged to ST241; PA521(36-5-29-7-4-10-7) belonged to ST170; PA536 (38-11-34-13-1-2-4) belonged to ST235; PA351 (5-1-9-40-1-16-24) belonged to ST238.

　　Conclusions:MLST is a highly discriminatory method which can be used to   standardize for. Pseudomonas aeruginosa typing. The result is unambiguous .The typing can help us to learn etiology, pathogenesis of the pathogen more further It also can give us a clue to understand the phylogenetic relationships and background between isolates better.

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