Detection of DNA derived from Trichinella spiralis by loop-mediated isothermal amplification

Abstract

Objective To detect Trichinella spiralis DNA by loop-mediated isothermal amplification (LAMP).Methods DNA was extracted by phenol-chloroform extraction method from Trichinella larva.Four primers were designed based on 18S rRNA sequence and used for LAMP assay.To evaluate the specificity of the LAMP method,DNA sample derived from Schistosoma japonicum was used as control.For evaluating the sensitivity of LAMP,the genomic DNA derived from T.spiralis was serially diluted and amplified by LAMP.LAMP reaction mixture was stained with SYBR green I dye and the result was determined according to the color developed in the tube by the naked eye.Green indicated positive reaction while brown was negative.Results After LAMP reaction,the test tube which used genomic DNA derived from T.spiralis appeared to be green,while control was brown.The sensitivity of LAMP assay in this study reached up to 415 fg/μl.Conclusion LAMP assay was established for the detection of DNA sample of T.spiralis. Key words: Trichinella spiralis ; Loop-mediated isothermal amplification ; Detection ;