CAJ | 학술논문

目的：建立16S rDNA PCR测序（PCR‐SBT）方法快速鉴定败血症患者体内的细菌菌种。方法对5株标准菌株及临床常见的20属27种细菌进行16S rDNA保守序列PCR扩增，扩增产物经纯化后直接测序，并利用Blast软件从GenBank数据库中搜索相关菌株的16S rDNA序列，采用Clustal‐X软件进行多序列比对和同源性分析，以确定细菌的种属；同时对124例败血症患者进行血培养并与PCR‐SBT 法比较，探讨其在临床应用中的价值。结果所有细菌菌株通过PCR方法均获得约1500 bp的保守序列，经测序分析后5株标准菌株的序列与预期标准序列完全一致，27种临床分离株测序鉴定结果与生化鉴定结果一致；124例败血症患者中84例PCR法阳性，阳性率67．7％，显著高于血培养的30．6％（P＜0．01）。结论建立16S rDNA的PCR‐SBT法鉴定细菌快速可靠，诊断败血症优于传统的血培养法。
목적：건립16S rDNA PCR측서（PCR‐SBT）방법쾌속감정패혈증환자체내적세균균충。방법대5주표준균주급림상상견적20속27충세균진행16S rDNA보수서렬PCR확증，확증산물경순화후직접측서，병이용Blast연건종GenBank수거고중수색상관균주적16S rDNA서렬，채용Clustal‐X연건진행다서렬비대화동원성분석，이학정세균적충속；동시대124례패혈증환자진행혈배양병여PCR‐SBT 법비교，탐토기재림상응용중적개치。결과소유세균균주통과PCR방법균획득약1500 bp적보수서렬，경측서분석후5주표준균주적서렬여예기표준서렬완전일치，27충림상분리주측서감정결과여생화감정결과일치；124례패혈증환자중84례PCR법양성，양성솔67．7％，현저고우혈배양적30．6％（P＜0．01）。결론건립16S rDNA적PCR‐SBT법감정세균쾌속가고，진단패혈증우우전통적혈배양법。
OBJECTIVE To establish 16S rDNA PCR sequence‐based typing (PCR‐SBT ) for the rapid identification of bacteria in the patients with sepsis .METHODS The 16S rDNA conservative sequence PCR amplification was performed for 5 standard strains and 27 species of bacteria of 20 common clinical genera ,the amplified products were directly sequenced after the purification ,the 16S rDNA sequences of the relevant strains were searched from the GenBank database by using Blast software ,the multiple sequence alignment and homological analysis were per‐formed with the use of Clustal‐X software so as to determine the species of the bacteria ,the blood culture was con‐ducted for 124 sepsis patients and was compared with the PCR‐SBT method ,and its clinical value was evaluated . RESULTS Totally about 1 500 bp conservative sequences have been acquired from all the strains through the PCR method ,and the sequencing analysis showed that the sequences of the 5 standard strains were completely consist‐ent with the expected standard strains ,and the results of the sequencing in identification of the 27 clinical isolates were as same as the results of the biochemical identification .Of the 124 sepsis patients ,84 were tested positive for the PCR method with the positive rate of 67 .7% ,significantly higher than 30 .6% of the blood culture (P<0 .01) .CONCLUSION The 16S rDNA PCR‐SBT is rapid and reliable for the identification of the bacteria and is su‐perior to the traditional blood culture method in the diagnosis of sepsis .