The LAMP method applied in the detection of carbapenem-resistance acinetobacter baumannii OXA-23 genes / 国际检验医学杂志

ABSTRACT

Objective To establish a simple ,rapid,highly specific and sensitive molecular detection of carbapenem-resistance acinetobacter baumannii(CRAB)OXA-23 genes,and this method is used to detect the multiple drug-resistant acinetobacter bau-mannii in our hospital,and the purpose is to know the antibiotic resistance of CRAB OXA-23 genes .Methods The loop-mediated isothermal amplification(LAMP)was established for detection of the CRAB OXA-23 genes,and a set of specific primers were de-signed by special software,PrimerExplorer version 4.The LAMP assay was developed on using SYBR Green Ⅰ for fluorescent chromogenic reaction substances,improved through a series of optimization tests,and through macroscopic observation and electro-phoresis test comparison results.At the same time,the application of LAMP was used to test 41 multiple drug-resistant acineto-bacter baumanniis which were collected from December 2013 to March 2014 in our hospitalized patients.Results The ladder ban-ding was produced in CRAB OXA-23 genes strains by the LAMP detection through electrophoresis test,however,no ladder ban-ding was observed in the others .The color of the amplification product in genes strain CRAB OXA-23 changed from orange to green by adding 1 μL SYBR Green Ⅰ,however it was still orange in others.The sensitivity of the LAMP detection in pure cultrue was 5 cfu/μL of the CRAB OXA-23 genes cells.Application of LAMP was used to separate multiple drug-resistant acinetobacter baumanniis from hospitalized patients ,32 strains were tested in 41 strains,the positive rate was 78.04%.Conclusion Separation of the CRAB OXA-23 genes carry rate is higher in our hospital ,and they have very high resistance of commonly used antibacterial drugs.The LAMP method to test OXA-23 gene of CRAB was established in this research was simple ,fast,sensitive and specific. Therefore,it is especially suitable wider use at the grass-roots unit,and it is of great significance for selecting reasonable choice of antibiotics by clinical doctor.